Inland recreational fisheries contribute nutritional benefits and economic value but are vulnerable to climate change.

Inland recreational fishing is primarily considered a leisure-driven activity in freshwaters, yet its harvest can contribute to food systems. Here we estimate that the harvest from inland recreational fishing equates to just over one-tenth of all reported inland fisheries catch globally. The estimated total consumptive use value of inland recreational fish destined for human consumption may reach US$9.95 billion annually. We identify Austria, Canada, Germany and Slovakia as countries above the third quantile for nutrition, economic value and climate vulnerability. These results have important implications for populations dependent on inland recreational fishing for food. Our findings can inform climate adaptation planning for inland recreational fisheries, particularly those not currently managed as food fisheries.

Identification of potential diagnostic and prognostic biomarkers for papillary thyroid microcarcinoma (PTMC) based on TMT-labeled LC-MS/MS and machine learning.

OBJECTIVES: To explore the molecular mechanisms underlying aggressive progression of papillary thyroid microcarcinoma and identify potential biomarkers. METHODS: Samples were collected and sequenced using tandem mass tag-labeled liquid chromatography-tandem mass spectrometry. Differentially expressed proteins (DEPs) were identified and further analyzed using Mfuzz and protein-protein interaction analysis (PPI). Parallel reaction monitoring (PRM) and immunohistochemistry (IHC) were performed to validate the DEPs. RESULTS: Five thousand, two hundred and three DEPs were identified and quantified from the tumor/normal comparison group or the N1/N0 comparison group. Mfuzz analysis showed that clusters of DEPs were enriched according to progressive status, followed by normal tissue, tumors without lymphatic metastases, and tumors with lymphatic metastases. Analysis of PPI revealed that DEPs interacted with and were enriched in the following metabolic pathways: apoptosis, tricarboxylic acid cycle, PI3K-Akt pathway, cholesterol metabolism, pyruvate metabolism, and thyroid hormone synthesis. In addition, 18 of the 20 target proteins were successfully validated with PRM and IHC in another 20 paired validation samples. Based on machine learning, the five proteins that showed the best performance in discriminating between tumor and normal nodules were PDLIM4, ANXA1, PKM, NPC2, and LMNA. FN1 performed well in discriminating between patients with lymph node metastases (N1) and N0 with an AUC of 0.690. Finally, five validated DEPs showed a potential prognostic role after examining The Cancer Genome Atlas database: FN1, IDH2, VDAC1, FABP4, and TG. Accordingly, a nomogram was constructed whose concordance index was 0.685 (confidence interval: 0.645-0.726). CONCLUSIONS: PDLIM4, ANXA1, PKM, NPC2, LMNA, and FN1 are potential diagnostic biomarkers. The five-protein nomogram could be a prognostic biomarker.

Priorities to inform research on marine plastic pollution in Southeast Asia.

Southeast Asia is considered to have some of the highest levels of marine plastic pollution in the world. It is therefore vitally important to increase our understanding of the impacts and risks of plastic pollution to marine ecosystems and the essential services they provide to support the development of mitigation measures in the region. An interdisciplinary, international network of experts (Australia, Indonesia, Ireland, Malaysia, the Philippines, Singapore, Thailand, the United Kingdom, and Vietnam) set a research agenda for marine plastic pollution in the region, synthesizing current knowledge and highlighting areas for further research in Southeast Asia. Using an inductive method, 21 research questions emerged under five non-predefined key themes, grouping them according to which: (1) characterise marine plastic pollution in Southeast Asia; (2) explore its movement and fate across the region; (3) describe the biological and chemical modifications marine plastic pollution undergoes; (4) detail its environmental, social, and economic impacts; and, finally, (5) target regional policies and possible solutions. Questions relating to these research priority areas highlight the importance of better understanding the fate of marine plastic pollution, its degradation, and the impacts and risks it can generate across communities and different ecosystem services. Knowledge of these aspects will help support actions which currently suffer from transboundary problems, lack of responsibility, and inaction to tackle the issue from its point source in the region. Being profoundly affected by marine plastic pollution, Southeast Asian countries provide an opportunity to test the effectiveness of innovative and socially inclusive changes in marine plastic governance, as well as both high and low-tech solutions, which can offer insights and actionable models to the rest of the world.

Evidence for causal effects of sleep disturbances on risk for osteoarthritis: a univariable and multivariable Mendelian randomization study.

OBJECTIVE: To disentangle whether sleep disturbances have a causal effect on the risk of osteoarthritis (OA) using genetically based approaches. METHOD: We performed univariable and multivariable Mendelian randomization (MR) analyses using publicly released genome-wide association studies summary statistics to estimate the causal associations of sleep disturbances with OA risk. The inverse-variance weighted (IVW) method was utilized as primary MR analysis, whereas complementary methods including weighted median, weighted mode, MR-Egger regression, and MR pleiotropy residual sum and outlier (MR-PRESSO) were applied to detect and correct for the presence of pleiotropy. RESULTS: There were 228 independent instrumental variables (IVs) for insomnia and 78, 27 and 8 IVs for sleep duration, short sleep duration and long sleep duration, respectively. Univariable MR analysis suggested that genetically determined insomnia or short sleep duration exerted a causal effect on overall OA in an unfavorable manner (Insomnia: OR = 1.22, 95%CI = 1.15-1.30, P = 8.05 × 10-10; Short sleep duration: OR = 1.04, 95%CI = 1.02-1.07, P = 2.20 × 10-3). More compelling, increasing genetic liability to insomnia or short sleep duration was also associated with OA risk, after accounting for effects of insomnia or short sleep duration on body mass index, type 2 diabetes and depression individually, and in a combined model considering all three confounders. CONCLUSIONS: Findings suggested consisted evidence for an adverse effect of increased insomnia or short sleep duration on OA risk. Strategies to mitigate sleep disturbances may be one of the cornerstones protects against OA.

Addressing the implementation challenge of the global biodiversity framework.

A Global Biodiversity Framework (GBF) is under discussion for the period 2021-2030, which will replace the "Aichi Targets" adopted by the Convention on Biological Diversity (CBD) in 2010. Given the limited success in meeting most of the Aichi Targets, this new framework must adopt a different approach. A key challenge the GBF must address is its implementation at national scales. Four ways this implementation challenge can be addressed include:The framework must move away from numerical targets to pursue positive trends in biodiversity, through adopting a "vectors of change" approach;The framework should be structured to focus on ecosystems and processes;The framework should synergise more extensively with existing biodiversity-relevant global agreements to maximise leverage and reduce overlap of resource use;The framework must adopt a much stronger theory of change than is in the current GBF Draft, to serve as the roadmap governments can use in upscaling their implementation of biodiversity conservation, sustainable use and benefit sharing. Finally, the GBF must become a "learning framework", committed to facilitating and enabling governments to each meet their specific biodiversity challenges, while sharing back experiences with the global community, leading ultimately to realising the 2050 CBD vision of people living in harmony with nature.

Arsenic enhances cell death and DNA damage induced by ultraviolet B exposure in mouse epidermal cells through the production of reactive oxygen species.

BACKGROUND: Ultraviolet (UV)B radiation has long been considered a carcinogen in both epidemiological surveys and experimental studies. However, recent work has suggested that different dosages of UVB exert different influences on cells. There are also co-carcinogenesis factors such as arsenic that affect the role of UVB. AIM: To explore the co-carcinogenesis effect of UVB and arsenic on the mouse epidermal cell line JB6 and the mechanism underlying it. METHODS: Growth of JB6 cells was measured by MTT assay. We carried out a comet assay to determine the DNA damage caused by UVB and arsenic, and tested the expression of DNA repair protein by western blotting. Reactive oxygen species (ROS) were measured using DCF and DHE staining, and changes in antioxidant enzymes were assessed using western blotting. RESULTS: Viability assays showed that arsenic increased the UVB-induced death rate. Arsenic enhanced DNA damage caused by UVB both directly by injury to double-stranded DNA and indirectly by reducing the capability of DNA repair in JB6 cells. All of these effects are the results of increased ROS generation and reduced expression of the antioxidant enzyme superoxide dismutase (SOD)1. CONCLUSION: Arsenic was found to enhance UVB-induced production of ROS and to downregulate SOD1 expression, leading to DNA damage and apoptosis in mouse skin cells. The combination of arsenic and UVB exposure was found to differentially regulate the expression of SOD1 and SOD2.

[A novel chemo-resistant gene MSX2 discovered by establishment of two pancreatic cancer drug resistant cell lines JF305/CDDP and PANC-1/GEM].

Objective: To explore new multidrug resistant genes of pancreatic cancer by establishment and characterization of chemo-resistant cell lines. Methods: The cisplatin-resistant cell line JF305/CDDP and the gemcitabine-resistant cell line PANC-1/GEM were induced by high-dose intermittent treatment. CCK-8 assay was used to detect the 50% inhibiting concentration (IC(50)), drug resistance index (R), cross-resistance, and growth difference of different cells. The changes of cell cycle and migration ability of drug-resistant cells were determined by flow cytometry and transwell assay, respectively. And then real-time fluorescence quantitative PCR was used to detect the expression of multidrug resistance-related genes. Results: The drug resistance indexes of JF305/CDDP and PANC-1/GEM were 15.3 and 27.31, respectively, and there was cross-resistance. Compared with the parental cells, the proliferation rate of JF305/CDDP was decreased by 40% on the fourth day (P<0.05); the proportion of S phase was decreased from (45±2)% to (30±2)% (P<0.05), and the migration ability was enhanced from (32 ±1) cells per field to (158±5) cells per field (P<0.01). The expression of multidrug resistance-related genes MRP2, MDR1, LRP and MSX2 was increased in JF305/CDDP cells (P<0.05). Knockdown of MSX2 in JF305 cells reduced the expression of MRP2, whereas overexpression of MSX2 in PANC-1 cells upregulated MRP2 level (P<0.05). Conclusions: Two stable multidrug resistant cell lines of pancreatic cancer, JF305/CDDP and PANC-1/GEM, were successfully established. MSX2 might be a new drug resistance related gene in pancreatic cancer cells by up-regulation of MRP2 expression.

Isotachophoretic Fluorescence in Situ Hybridization of Intact Bacterial Cells.

A counter-pressure-assisted capillary isotachophoresis method in combination with a sieving matrix and ionic spacer was used to perform in-line fluorescence in situ hybridization (FISH) of bacterial cells. A high concentration of sieving matrix (1.8% w/v HEC) was introduced at one end of the capillary, and the bacterial cells were suspended in the spacer electrolyte for injection. Using a 2 min injection with 18 psi counter-pressure, 50% of the cells injected into the capillary were hybridized with the fluorescently labeled oligonucleotide, and the excess unhybridized probe was separated from the hybridized cell-probe complexes in a two-stage ITP method. With an LOD (6.0 × 104 cells/mL) comparable with the CE analysis of a sample processed using an off-line FISH protocol, the total analysis time was reduced from 2.5 h to 30 min. Provided the appropriate probe is selected, this approach can be used for specific detection of bacterial cells in aqueous samples.

Fibre-based electrofluidics on low cost versatile 3D printed platforms for solute delivery, separations and diagnostics; from small molecules to intact cells.

A novel and effective fibre-based microfluidic methodology was developed to move and isolate charged solutes, biomolecules, and intact bacterial cells, based upon a novel multi-functional 3D printed supporting platform, with potential applications in the fields of microfluidics and biodiagnostics. Various on-fibre electrophoretic techniques are demonstrated to separate, pre-concentrate, move, split, or cut and collect the isolated zones of target solutes, including proteins and live bacterial cells. The use of knotting to link different fibre materials, and the unique ability of this approach to physically concentrate solutes in different locations are shown such that the concentrated solutes can be physically isolated and easily transferred to other fibres. Application of this novel fibre-based technique within a potential diagnostic platform for urinary tract infection is shown, together with the post-electrophoretic incubation of live bacterial cells, demonstrating the cell survival following on-fibre electrophoretic concentration.

Stereotactic radiosurgery for the treatment of mesial temporal lobe epilepsy.

OBJECTIVES: Stereotactic radiosurgery (RS) is a potential option for some patients with temporal lobe epilepsy (TLE). The aim of this meta-analysis was to determine the pooled seizure-free rate and the time interval to seizure cessation in patients with lesions in the mesial temporal lobe, and who were eligible for either stereotactic or gamma knife RS. MATERIALS & METHODS: We searched the Medline, Cochrane, EMBASE, and Google Scholar databases using combinations of the following terms: RS, stereotactic radiosurgery, gamma knife, and TLE. RESULTS: We screened 103 articles and selected 13 for inclusion in the meta-analysis. Significant study heterogeneity was detected; however, the included studies displayed an acceptable level of quality. We show that approximately half of the patients were seizure free over a follow-up period that ranged from 6 months to 9 years [pooled estimate: 50.9% (95% confidence interval: 0.381-0.636)], with an average of 14 months to seizure cessation [pooled estimate: 14.08 months (95% confidence interval: 11.95-12.22 months)]. Nine of 13 included studies reported data for adverse events (AEs), which included visual field deficits and headache (the two most common AEs), verbal memory impairment, psychosis, psychogenic non-epileptic seizures, and dysphasia. Patients in the individual studies experienced AEs at rates that ranged from 8%, for non-epileptic seizures, to 85%, for headache. CONCLUSION: Our findings indicate that RS may have similar or slightly less efficacy in some patients compared with invasive surgery. Randomized controlled trials of both treatment regimens should be undertaken to generate an evidence base for patient decision-making.

Anti-prostate cancer effects of CTL cell induction in vitro by recombinant adenovirus mediated PSMA/4-1BBL dendritic cells: an immunotherapy study.

This study aimed to examine anti-prostate cancer immune response induced by dendritic cells (DCs) transduced with PSMA/4-1BBL recombinant adenoviruses in vitro. Ad-PSMA, Ad-4-1BBL, and Ad-GFP were transfected into DCs derived from peripheral blood of healthy volunteers. Ad-PSMA/4-1BBL-DC, Ad-PSMA-DC, Ad-4-1BBL-DC, Ad-GFP-DC, and normal-DC, PSMA and 4-1BBL protein levels in DCs were detected by western blot. IL-12, IFN-γ and IL-10 were measured by ELISA. Mixed lymphocyte reaction and the cytotoxicity of each group targeted to LNCap, Du145, and 22RV prostate cancer cells were determined by CCK-8 assay. PSMA and 4-1BBL protein could express on DC successfully, the IL-12 supernatant content (134.29 ± 2.22 pg) was higher than others (P < 0.05). The ability to stimulate autologous T lymphocyte proliferation in the co-transfection group was higher than others (P < 0.05). When the DCs were co-cultured with CTLs, the PSMA/4-1BBL-DC-CTL group showed the highest content of IFN-γ (1176.10 ± 14.37pg/5 x 10(6) cells), but the lowest IL-10 content (75.14 ± 2.01 pg/5 x 10(6) cells) (P < 0.05), and the strongest anti-tumor effect when the effector to target ratio was 40:1, along with a higher killing ratio of LNCap cells than others (P < 0.05). Overall, Mature DCs transfected with Ad-PSMA/4- 1BBL not only showed high secretion of IL-12, but also induced CTLs to stimulate and enhance the killing effect of PSMA specific effector cells to PSMA positively expressing prostate cancer cells. Furthermore, the DCs infected with two kinds of tumor-associated antigens would induce more effective tumor-specific CTL induction.

The effect of mahogunin gene mutant on reproduction in male mice: a new sight for infertility?

Mahogunin is an important mediator of chromogenesis and neurodegeneration. Mahoganoid is a mutation of the mahogunin gene, which causes a pleiotropic phenotype that includes suppression of obesity, spongiform neurodegeneration and improvement of insulin sensitivity. Our previous research found that mahoganoid widely expressed in the male rat reproductive system, and mahoganoid-deficient mice have reduced embryonic viability. But the reproductive change in mahogunin knockout (md(nc) ) male mice has not been reported previously. Here, we report that the mahogunin mRNA also widely exists in reproductive system of male mice, and its mRNA expression in the testis was in accordance with the first spermatogenesis wave cycle. Moreover, we find that md(nc) male mice were able to mate with females but no pups are delivered. Besides, the sperms' active progressive motility and hormone secretion (E2, FSH, LH, PRL) were obviously decreased while abnormal sperm rate showed no significant difference in md(nc) compared to wild-type (WT) male mice. This study indicates the mahogunin deficiency results in the infertility of male mice, disruption of hormones secretion and impaired active progressive motility, which may additionally illuminate the aetiology of male infertility in human.

microRNA-199a-5p protects hepatocytes from bile acid-induced sustained endoplasmic reticulum stress.

Sustained endoplasmic reticulum (ER) stress has been linked to cell death and the pathogenesis of many liver diseases, including toxic liver, cholestasis, and infectious liver disease. The cellular pathways that attenuate hepatic ER stress have been the focus of many recent studies, but the role of microRNAs (miRNA) in this process remains unknown. Here, we report that one of the most abundant miRNAs in hepatocytes, miR-199a-5p, was elevated in both bile acid- and thapsigargin (TG)-stimulated cultured hepatocytes, as well as in the liver of bile duct-ligated mice. We identify the misfolded protein chaperone GRP78, as well as the unfolded protein response transducers endoplasmic reticulum to nucleus signaling 1 and activating transcription factor 6 as direct targets of miR-199a-5p, and show that endogenous miR-199a-5p represses the 3' untranslated regions (UTRs) of their mRNAs. Through gain-of-function and loss of function approaches, we demonstrate that the elevated miR-199-5p disrupts sustained ER stress and prevents hepatocytes from undergoing bile acid- or TG-induced cell death. Furthermore, we reveal that the transcription factor AP-1 is a strong positive regulator of miR-199a-5p. In brief, our study demonstrates that AP-1/miR-199a-5p and ER stress mediators form a feedback loop, which shields hepatocytes from sustained ER stress and protects the liver from injury. On the basis of these findings, we also suggest that the miRNA miR-199a-5p is a potential target for clinical approaches aiming to protect hepatocytes in liver disease.

First Report of Curvularia Leaf Blight on Curcuma wenyujin Caused by Curvularia clavata in China.

Curcuma wenyujin Y.H. Chen & C. Ling is a traditional Chinese medicinal herb in the Zingiberaceae family. Commonly known as Wen yujin, the root is widely used for alleviating pain and protecting the liver. A severe leaf blight disease was observed in three C. wenyujin farms in Hainan Province of China in October 2010. The obvious symptoms of leaf blight, yellow to brown irregular lesions (1 to 20 cm) on C. wenyujin, usually began at the tips of leaves and the main veins. This disease, especially severe from August to October, caused heavy damage and 100% of mature plants (10 months old) in farms were infected. The disease was most severe when continuous cropping was performed and showed slight improvement when rotation was adopted. Farmers usually sprayed carbendazim (50% WP) and thiophanate-methyl (70% WP) to control this disease, but these treatments were not effective. To isolate the causal pathogen, diseased plants were collected in October 2010 from a field of the Hainan Branch Institute of Medicinal Plant Development in Hainan Province. Lesion tissue was removed from the border between symptomatic and healthy tissue, surface sterilized in 75% ethanol for 1 min, washed in three changes of sterile distilled water, transferred to potato dextrose agar (PDA) plates, and incubated at 28°C for 7 days. Single spore cultures of five isolates were obtained and identified as Curvularia clavata based on morphological characteristics (1). Conidia measured 20 to 29 × 7.5 to 10.5 µm (n = 100), were curved, 3-septate, and the third cell from the base was larger and darker than the others. Mycelia of single spore cultures growing on PDA for 5 days were used for DNA extraction using a plant genomic DNA kit (TIANGEN, Beijing). The internal transcribed spacer (ITS) region of the rDNA was amplified using primers ITS1 and ITS4. The amplicons were 562 bp in length (GenBank Accession No. JQ730852) and had 99% nucleotide identity with the GenBank Accession No. JN021115 and AF071336 of C. clavata. Pathogenicity tests were conducted using fresh and healthy detached Curcuma wenyujin leaves. Mycelial discs (10 mm) removed from a 5-day-old colony on PDA were used for inoculation. Each isolate was inoculated on three distinct leaves (two distinct inoculations per leaf). Three additional leaves inoculated with sterile PDA discs were used as control. Inoculated leaves were covered with a polythene film to maintain high humidity. Leaves in trays were kept in a growth chamber at 28°C and observed for symptom appearance every day. Five days after inoculation, inoculated leaves developed blight symptoms similar to those observed on naturally infected leaves. No symptoms were observed on non-inoculated leaves. C. clavata was reisolated from the inoculated leaves, thus fulfilling Koch's postulates. C. clavata has been previously reported to be economically important on a number of other hosts (2). To our knowledge, this is the first report of Curvularia leaf blight on Curcuma wenyujin caused by C. clavata in China. References: (1) A. M Mandokhot et al. Eur. J. Plant Pathol.78:65, 1972. (2) T. Y. Zhang et al. Flora fungorum sinicorum: Beijing, China, 2010.

Adjuvant transcatheter arterial chemoembolization for intrahepatic cholangiocarcinoma after curative surgery: retrospective control study.

BACKGROUND: Effects of adjuvant transcatheter arterial chemoembolization (TACE) for intrahepatic cholangiocarcinoma (ICC) radical surgery have never been evaluated. METHODS: A retrospective analysis was conducted on 125 ICC patients who had undergone operations with curative intent in Shanghai Eastern Hepatobiliary Surgery Hospital from July 2002 to December 2003. Of these patients, 53 underwent adjuvant TACE (TACE group) and 72 did not (non-TACE group). Adjuvant TACE was performed one time 1.5-2.0 months after the operation. RESULTS: Follow-up was performed at a median of 18 months (range 3-96 months). There was no significant recurrence-free survival (RFS) difference between the TACE and non-TACE groups (P = 0.659). The 1-, 3-, and 5-year overall survival (OS) rates were 69.8, 37.7, and 28.3%, respectively, for the TACE group and 54.2, 25.0, and 20.8%, respectively, for the non-TACE group (P = 0.045). Among 54 patients with a recurrence time of ≤ 3 months, the OS rate of the TACE group was better than that of the non-TACE group (P < 0.001). For 59 patients with a recurrence time later than the median RFS, no significant RFS difference was found between the TACE and non-TACE groups (P = 0.681). These results indicate that TACE could not delay recurrence but could prolong the OS of patients with early recurrence. CONCLUSIONS: Adjuvant TACE after radical surgery was associated with better survival among the ICC patients with early recurrence.

First Report of Corynespora Leaf Spot on Patchouli Caused by Corynespora cassiicola in China.

Patchouli (Pogostemon cablin (Blanco) Benth.) is mainly cultivated in Southeast Asia as a medicinal shrub and a source of patchouli oil used in perfumery. In 2008, a leaf spot disease was observed on patchouli plants grown on most farms (some farms had 99% incidence) in Wanning, the predominant cultivation location in the Hainan Province of China. The disease usually began at the tip of leaves, the main veins, or small veinlets. Severely irregular-shaped dark brown leaf spots expanded over 5 to 10 days, eventually causing infected leaves to abscise. The time from initial leaf lesions to abscission usually took 1 month. The disease was usually most severe in April and May, causing significant economic losses along with quality losses to patchouli oil extracted from leaves. To isolate the causal pathogen, diseased leaves were collected in August 2008 from a farm of the Hainan Branch Institute of Medicinal Plant Development in Wanning, surface sterilized in 75% ethanol for 1 min, transferred to potato dextrose agar (PDA), and incubated at 28°C for 14 days. Single-spore cultures of three isolates were obtained and identified as Corynespora cassiicola (Berk. & Curt.) Wei. on the basis of morphological and physiological features (1). Genomic DNA was extracted from all the cultures. The internal transcribed spacer (ITS) region of the rDNA was amplified using primers ITS1 (5'-TCCGATGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). Amplicons were 546 bp (GenBank Accession No. HM145960) and had 99% nucleotide identity with the corresponding sequence (GenBank Accession No. GU138988) of C. cassiicola isolated from cassava (Manihot esculenta Crantz). To satisfy Koch's postulates, 50-day-old potted plants in a tent were sprayed until runoff with a spore suspension (1 × 106 spores/ml) prepared from 10-day-old cultures. Using this spray method, one isolate was inoculated separately onto nine leaves of three potted plants. The potted plants were covered with plastic bags to maintain high humidity for 48 h and then placed outside under natural environmental conditions (temperature 20 to 28°C). Another nine leaves of three potted plants, sprayed only with sterile water, served as noninoculated control plants. Leaf spot symptoms similar to those on diseased field plants appeared after 7 days on all inoculated plants. C. cassiicola was reisolated from all inoculated test plants. No symptoms were observed on the control plants. To our knowledge, this is the first report of C. cassiicola causing a leaf spot disease on patchouli in China. Other previous reports of this disease were from Cuba (2). This pathogen has also been reported previously to be economically important on a number of other hosts. On patchouli plants, more attention should be given to prevention and control measures to help manage this disease. References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute: Kew, Surrey, England, 1971. (2) I. Sandoval et al. Cienc. Tec. Agric., Prot. Plant. 10:21, 1987.

First Report of Alternaria Leaf Blight on Bupleurum chinense Caused by Alternaria alternata in China.

Bupleurum chinense DC. (family Umbelliferae) is an important medicinal herb in traditional Chinese medicine and is cultivated as an economically important plant in China (2). From 2006 to 2009, severe foliar disease was observed on B. chinense in Haidian, Changping, and Shunyi districts, Beijing, China. Approximately 75 to 85% of fields were affected with disease incidence ranging from 65 to 90%. Distribution of the disease in affected B. chinense fields was generally associated with high soil moisture, often corresponding to poor drainage. Initial symptoms first appeared on older leaves as irregularly shaped, minute, dark brown-to-black spots, with yellow borders on the edge of the affected leaflet blade. As the disease progressed, the lesions expanded, causing the leaflets to turn brown, shrivel, and die. Isolations performed on potato dextrose agar (PDA) initially resulted in white colonies. After 7 days of incubation at 25°C, the colonies turned gray or brown. Conidia varied in size from 10 × 6 to 40 × 12 µm, appeared brown to dark brown or olive-brown, were short beaked and borne in long chains, oval and bean-shaped with one to six transverse septa and zero to three longitudinal septa. Sequences of the rDNA from the internal transcribed spacer regions 1 and 2 and the 5.8S gene were amplified using primers ITS1 and ITS4, were obtained from three isolates, and comparisons with GenBank showed 100% similarity with A. alternata (Genbank Accession No. AB470912.1). For pathogenicity tests, three isolates were grown on PDA for 14 days. Inoculations were performed on detached, surface-sterilized, and healthy B. chinense leaflets following the method of Belisario (1). A 5-µl drop of conidial suspension containing 1 × 105 CFU/ml was placed on each leaflet and 12 leaves per isolate were used. Leaves were incubated in a growth chamber (80 to 90% relative humidity; 50 to 60 klx/m2 light intensity with a 12-h photoperiod). After 5 days, leaf spots similar to the original symptoms developed on all inoculated leaves and A. alternata was consistently reisolated from symptomatic leaf tissues on PDA. Control leaflets inoculated with sterile water remained asymptomatic. The experiment was performed three times. To our knowledge, this is the first report of A. alternata on B. chinense from China. References: (1) A. Belisario et al. Plant Dis. 83:696, 1999. (2) C. Sui et al. Plant Dis. 93:844, 2009.

First Report of Broad bean wilt virus 2 Infecting Bupleurum chinense in China.

Samples of the medicinal plant, Bupleurum chinense DC., were collected in October 2007 from the garden of the Institute of Medicinal Plant Development in Beijing. Partial fragments of the genomic RNA1 and RNA2 of Broad bean wilt virus 2 (BBWV-2) were obtained from the root cDNA library of these plants. Sequence analyses showed the 1,896-nt RNA1 fragment (GenBank No. FJ485684) encoding a portion of the RNA-dependent RNA polymerase (RdRp) and the 2,017-nt RNA2 fragment (No. FJ485685) encoding 612 amino acids of the complete large (LCP) and small coat protein (SCP), respectively. The amino acid identities of LCP and SCP were 90.8 to 96.7% compared with sequences of other BBWV-2 isolates deposited in the GenBank with the highest homology to Japanese IP (No. AB018698) and the lowest to Japanese 1-2 (No. AB018701). This strongly suggests that the B. chinense plants utilized for cDNA library construction were infected by what appears to be an isolate of BBWV-2. Seeds from the same batch were sown again in the same garden in May 2008. In August 2008, approximately 30% of these plants showed mosaic, distortion, and stunting. Reverse transcription (RT)-PCR amplicons were obtained from eight symptomatic plants using a pair of conserved primers for specific detection of viruses within the Fabavirus genus (2). A symptomless plant tested negative by RT-PCR. The same single 391-bp amplicon of RNA1 (No. FJ485686) obtained from five of those eight symptomatic plants were cloned and sequenced. Sequence comparison with the corresponding sequences of other BBWV-2 isolates showed that the sequenced isolate was most closely related to B935-a Chinese faba bean isolate (No. AF149425). Crude sap of one diseased B. chinense plant was used for mechanical inoculation to Chenopodium amaranticolor Coste & Reyn. Chlorotic local lesions were observed on inoculated leaves 5 days after inoculation, and subsequently, systemic mottle and malformed symptoms appeared on the upper leaves. Twelve plants were inoculated and all plants showed symptoms of virus infection. RT-PCR tests of inoculated indicator plants showing local lesions confirmed the presence of BBWV-2. To date, Clover yellow vein virus and Lettuce mosaic virus have been isolated from the genus Bupleurum (B. griffithii hort. and B. falcatum L. sensu lato) in Japan and Israel, respectively (1,3). Furthermore, to our knowledge, no genomic sequence of BBWV-2 naturally infecting plants in the family Umbelliferae/Apiaceae has been reported. Therefore, this is the first report of BBWV-2 on B. chinense (Umbelliferae/Apiaceae), which was designated as a BC isolate of BBWV-2. In China, BBWV-2 was reported to be infecting and causing heavy losses to many plant species mostly belonging to the family Leguminosae (4). B. chinense is a commonly used bulk medicinal plant mainly cultivated in Hebei, Sichuan, Gansu, and Shanxi provinces in China for decoction pieces and extracts of its dried roots, which are also exported to Japan, Korea, and Southeast Asia. These results demonstrate the need for further assessment of BBWV-2 incidence and the losses it may cause. References: (1) J. Cohen et al. Phytoparasitica 30:88, 2002. (2) R. M. Ferrer et al. J. Virol. Methods 144:156, 2007. (3) H. Yamamoto. Jpn. J. Phytopathol. 69:420, 2003. (4) X. P. Zhou et al. Acta Phytopathol. Sin. 26:347, 1996.

Expression of HSP70 and JNK-related proteins in human liver cancer: Potential effects on clinical outcome.

BACKGROUND: Activation of stress-activated protein kinase/c-Jun N-terminal kinase was inhibited in cells, in which heat shock protein70 was induced to a high level, indicating that heat shock protein70 might be anti-apoptosis protein. AIM: We examined the expression of heat shock protein70 and c-Jun N-terminal kinase signal transduction pathway in human liver carcinoma to explore their relationship and clinical parameters. PATIENTS AND METHODS: The expression of heat shock protein70, c-Jun N-terminal kinase1, c-Jun N-terminal kinase2 and c-Jun were detected immunohistochemically in 62 samples of liver cancer. Western blot was used to confirm immunostaining results. RESULTS: Heat shock protein70 expression showed a positive correlation with the malignant differentiation in liver carcinoma (r=0.449, P<0.0005). The expression of c-Jun N-terminal kinase1, c-Jun N-terminal kinase2, and c-Jun showed a negative correlation with the malignant differentiation in liver carcinoma (r=-0.351, P=0.005; r=-0.303, P=0.017; r=-0.302, P=0.017). Heat shock protein70 expression was correlated with c-Jun N-terminal kinase1 (r=-0.385, P=0.002), c-Jun N-terminal kinase2 (r=-0.309, P=0.015) and c-Jun (r=-0.302, P=0.017). Expression of heat shock protein70, as well as c-Jun N-terminal kinase1, was correlated with recurrence-free survival after the resection. Heat shock protein70 was associated with prognosis (P=0.004). CONCLUSION: Expression of heat shock protein70 and c-Jun N-terminal kinase-related proteins might be an indicator of malignant potential in liver carcinoma. The balance between heat shock protein70 and c-Jun N-terminal kinase-related protein may increase the stability of liver cancer cells in stress. Negative expression of heat shock protein70 might be a protective factor of recurrence of liver carcinoma.