First report of Brown blotch disease caused by Diaporthe ueckeri on Hyophorbe lagenicaulis in China.

Bottle palm (Hyophorbe lagenicaulis) is a picturesque evergreen plant in the family Aceraceae, ubiquitously cultivated as an ornamental tree throughout the tropics and subtropics due its attractive shape and small stature. During 2016-2022, brown spots were observed on the leaves of bottle palm on both sides of a campus road in Mazhang, Zhanjiang, Guangdong province (N21°8' 59.9" E110°17' 51.4"). Symptoms appeared as circular, light yellow to brownish red, slightly sunken spots, and brown to black in the center (Fig 1 A-C). The spots expanded to big irregular blotches, which finally led leaves to wither. 0ne hundred percent of 50 plants were infected and 90% of the leaves each plant were covered with brown patches of different sizes. Tissues (5 × 5 mm) from the diseased-healthy junction of the leaf spots were surface disinfected with 75% ethanol for 30 s and 3.5% hydrogen peroxide solution for 5 min, rinsed three times with sterile water and placed on potato dextrose agar (PDA) at 25-28℃ in the dark for 3 days. Fungi with the similar morphology grew from 100% of these inoculated tissues. Hyphal tips from the colony edges were transferred to PDA. Single isolates were obtained by plate dilution method after sporogenesis. Colonies with sparse aerial mycelium were flat, grey. Conidiomata were superficial, black, solitary, scattered. Conidiophores were cylindrical, densely aggregated. Alpha conidia with bi-multiguttulate were hyaline, smooth, ellipsoidal, aseptate, 4.7-7.9×1.6-3.4 µm (Av. 6.4×2.3µm, n>50). Beta conidia were filiform, slightly curved, 15.9-28.3× 0.7-1.1 µm (20.1 × 0.9 µm, n>50) (Fig 1 E-H). Gamma conidia were not observed. The internal transcribed spacer (ITS) regions, large subunit (LSU) rRNA sequence, translation elongation factor (TEF), actin (ACT), calmodulin (CAL), histone H3 (HIS), and beta tubulin (TUB2) genes were amplified using the primers ITS4/ITS1 (White et al. 1990), LR0R/LR5, EF1-728F/EF-2, ACT-512F/ACT-783R, CAL228F/ CAL737R, CYLH3F/ H3-1b, and TUB2Fd /TUB4Rd (Aveskamp et al. 2009; Carbone and Kohn 1999; Crous et al. 2004; O'Donnell et al. 2000), respectively. All seven sequences of the isolate CCAS-JPYZ-4B (ACCC 35493) were submitted to NCBI (OR430112-3, OR451702-6). BLAST search result showed high sequence identity with several Diaporthe ueckeri isolates (Tab1). Phylogenetic analysis of the concatenated data of CAL, HIS, ITS, TEF and TUB genes using Maximum Likelihood method placed CCAS-JPYZ-4B in the D. ueckeri clade. Thus, the isolate was identified as D. ueckeri (Udayanga et al. 2015). Pathogenicity tests were performed on three 5-year-old plants, one healthy new leaf per plant and 10 sites on per leaf were slightly wounded and inoculated with 5-mm mycelial plugs from 10-day-old culture on PDA. The control sites were treated with PDA plugs. Each inoculated leaf was covered with a plastic bag to maintain high humidity and kept at natural temperatures (25-28 ℃). The experiment was repeated once. Symptoms appeared as those described as above 5 to 10 days after inoculation (Fig 1 D). Controls were asymptomatic. The fungus was reisolated from diseased leaves and identified as D. ueckeri. D. ueckeri may infect Arachis hypogaea, Cucumis melo, Camellia sinensis, Glycine max, Mangifera indica and Michelia shiluensis, and this is first report causing brown blotch on bottle palm in China. This disease occurred all year round, which seriously affected plants growth and ornamental value; it is necessary to develop an effective management strategy.

Erysiphe elevata causing powdery mildew on Eucalyptus urophylla × E. camaldulensis in China.

Eucalyptus urophylla × E. camaldulensis, named Chiwei eucalypt, is a hybrid species widely used in China. Many of its clones are cultivated for afforestation due to cold tolerance, high yield, high strength and disease resistance. Clone LH1 is planted extensively for its high stability and machinability in South China. In December 2021, severe powdery mildew signs were observed on clone LH1 in Zhanjiang, Guangdong (N28°8'29"; E110°17'5"). Whitish powder principally appeared on both abaxial and adaxial leaf surfaces. All plants were infected within about a week and above 90% leaves were diseased, which resulted in abnormal growth and shrinkage of leaves. Hyphae with single, lobed appressoria were hyaline, septate, branched, 3.3-6.8 µm (ave. 4.9 µm, n>50) wide. Conidiophores with a straight to flexuous foot-cell (14.7-46.1×5.4-9.7 µm, ave. 25.8×7.9 µm, n>30) were erect, hyaline, 2-septate, unbranched, 35.4-81.8 × 5.7-10.7 (ave. 56.7×8.7 µm, n>50). Conidia were solitary, hyaline, cylindrical to elliptical, 27.7-46.6 ×11.2-19.0 (ave. 35.7×16.6 µm, n>50). Chamothecia were not found on infected trees. The further identification was confirmed by partial sequences of internal transcribed spacer (ITS), large submit rRNA gene (LSU), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine synthetase (GS), and RNA polymerase II second largest subunit (RPB2) gene. A very small amount of mycelia and spores from voucher specimens CCAS-ASBF-1 and CCAS-ASBF-2 were deposited in the herbarium of Guangdong Ocean University. Specimens were PCR amplified and sequenced with primer pairs ITS1/ITS4 (White et al 1990), LROR/LR7 (Moncalvo et al 1995), PMGAPDH1/PMGAPDH3R, GSPM2/GSPM3R and PmRpb2_4/ PMRpb2_6R (Bradshaw, et al. 2022), respectively. BLASTn results showed that ITS (OP270019 and OQ380937), LSU (OP270018 and OQ380938), GAPDH, GS and RPB2 (OQ414445- OQ414450) were above 99% identical with those of E. elevata on Catalpa bignonioides (ITS: AY587013) (Cook et al 2004), Plumeria rubra (ITS: MH985631) (Yeh et al 2019), Cerbera manghas (ITS: MZ379159; LSU: MZ379160) (Mukhtar et al 2022), and Eucalyptus camaldulensis (LSU: LC177375-6) (Meebon et al. 2017), and above 99% identical with those of Erysiphe vaccinii FH00941201 on Vaccinium corymbosum (ITS: ON073869; RPB2: ON119159; GS: ON075687) and FH00112205 on V. vacillans (ITS: ON073870; GAPDH: ON075646) (Bradshaw et al 2022). This is the first sequence data for non rDNA of E. elevata. In an ITS tree phylogenetic analysis with Maximum likelihood (ML) method showed the fungus clustered in a highly supported clade with E. elevata and E. vaccinii. In a multi-locus tree, E. elevata grouped in a sister position to E. vaccinii FH00941201. Thus, the pathogen was identified as E. elevata based on morphology, DNA BLASTn and phylogenetic analysis (Braun and Cook 2012). Pathogenicity tests were conducted on healthy leaves of 1-year-old potted plants. Ten leaves were cleaned with sterile water, inoculated by gently dusting conidia from single lesion on the naturally infected leaves, and then covered with plastic bags containing wet absorbent cotton. Non-inoculated leaves served as controls. Symptoms developed on all inoculated leaves 3 to 5 days after inoculation, and the fungus was identical to the original fungus on the infected leaves, whereas control plants remained symptomless. This is the first report of powdery mildew caused by E. elevata on Eucalyptus sp. from China. This finding is helpful for land managers to diagnose and control the disease.

Associations of the Gut Microbiota Composition and Fecal Short-Chain Fatty Acids with Leukocyte Telomere Length in Children Aged 6 to 9 Years in Guangzhou, China: A Cross-sectional Study.

BACKGROUND: Telomere length (TL) serves as a marker of cellular senescence and appears to plateau between the age of 4 y and young adulthood, after which the gut microbiota are supposed to be established. However, scarce data are available regarding the correlation between gut microbiota composition and TL in the pediatric population. OBJECTIVES: We aimed to investigate whether the gut microbiota and the concentrations of SCFAs in feces are associated with leukocyte TL in children. METHODS: In total, 401 children aged 6-9 y from Guangzhou were enrolled in this cross-sectional study. qPCR was used to determine relative TL in peripheral blood leukocytes. The gut microbiota was characterized by 16S ribosomal RNA amplicon sequencing and the fecal concentrations of total SCFAs and SCFA subtypes were determined using HPLC. The multivariate methods with an unbiased variable selection (MUVR) algorithm and partial least square models were used to select predictable operational taxonomic units (OTUs). Further correlation analyses were performed based on multiple linear regression models with adjustment for covariates and false discovery rate. RESULTS: With the use of MUVR, 35 relevant and minimal optimal OTUs were finally selected. Multiple linear regression analysis showed that the abundance of several OTUs, including OTU334 (belonging to the genus Family XIII AD3011 group), OTU726 (belonging to the species Lachnoclostridium phocaeense), OTU1441 (belonging to the genus Ruminococcus torques group), OTU2553 (belonging to the genus Lachnospiraceae UCG-010), and OTU3375 (belonging to the family Lachnospiraceae), was negatively associated with leukocyte TL (ß: -0.187 to -0.142; false discovery rate (FDR)-corrected P value (PFDR) = 0.009-0.035]. However, neither SCFA subtype nor total SCFA content in feces exhibited significant associations with TL (ß: -0.032 to 0.048; PFDR = 0.915-0.969). CONCLUSIONS: The gut microbiota, but not fecal SCFA concentration, was significantly associated with TL in this pediatric population.

Leaf Spots on bodhi tree (Ficus religiosa) caused by Diaporthe tulliensis.

Ficus religiosa L., known as bodhi tree, is an ornamental tree and widely planted as an avenue and roadside tree due to ovate-rounded leaves with narrow, elongated tips. During 2018-2021, circular to oval-shaped leaf spots with pale white centers and brown-black edges surrounded by a chlorotic halo were observed on the leaves of more than 200 bodhi trees all year round in a park in Zhanjiang, Guangdong (N 21°15'22.29''; E110°23'1.03''). The leaf disease incidences were usually 15-80%, in severe cases, up to 100% in autumn and winter every year, and some trees shed all leaves(Fig S1). Repeated annual defoliation may weaken the tree and decrease the aesthetic value in the landscape. Diseased tissues (5 × 5 mm) of five symptomatic infected leaves were surface sterilized in 3% hydrogen peroxide solution for 3 min, rinsed thrice with sterile water, plated on potato dextrose agar (PDA) amended with ampicillin (50mg/L), and incubated at 25-28 ℃ in the dark for 3-7 days. Five strains with similar morphology were obtained by transferring hyphal tips of the colonies to fresh PDA and further isolating by single spore method. Fungal colonies were flat and spreading, with sparse, white aerial mycelium, and black pycnidial conidiomata semi-immersed in PDA after 30-days incubation at 25-28 ℃ in dark. Conidiophores were hyaline and α-conidia were single-celled, oval to fusiform, guttulate, 5.3 × 2.5 µm (n>50), similar to Diaporthe sp. (Crous et al. 2015), but no ß and γ -conidia were observed. The internal transcribed spacer(ITS), large subunit ribosomal RNA gene(LSU), calmodulin (CAL) and ß-tubulin(TUB) gene regions of a representative strain were amplified using specific primers reported by White et al. (1990), Gao et al (2017) and Gomes et al (2013), and submitted to GenBank (ITS: OM200852, LSU: OM228732, CAL: OM244761, TUB: OM244760). NCBI Blastn showed above 99% identity to D. tulliensis (anamorph: Phomopsis heveicola ) isolates of ITS (MT974186, MN393590 and KX457967), LSU (KR936131), CAL (MW759801), and TUB (KR936132 and MN399886), respectively (Crous et al. 2015; Huang et al. 2021; Gong et al. 2020; Bai et al. 2017). Based on the concatenated ITS, CAL, and TUB, a Maximum Likelihood phylogenetic analyses using MEGA 10.1.8 clustered the fungus with D. tulliensis in a clade with a 93% bootstrap support(Fig S2). Therefore, the fungus was identified as D. tulliensis based on morpho- molecular characteristics. Healthy detached leaves were sanitized thrice with 70% alcohol, and rinsed with sterile water. PDA plugs with actively growing 10-days-old mycelium were placed on predetermined sites, put into a sealed box with above 80% relative humidity and incubated at room temperature (25-28℃). Each isolate was inoculated at 25 needle-wounded and unwounded sites, PDA plugs without mycelium served as controls. Symptomatic spots appeared on all wounded leaves by 7 days post-inoculation (dpi) and on all unwounded leaves by 12 dpi. No symptoms appeared on controlled leaves. Pure cultures were recovered from inoculated leaves and showed identical morpho-molecular criteria to the original isolates. More than 70 pathogenic fungi are known to cause diseases on F. religiosa while there is no record of D. tulliensis infecting bodhi according to the U.S. National Fungus Collection (Farr and Rossman 2022). This report could provide basic understanding and alerting role for horticulturist in daily management.

[Neuropsychological development of late preterm infants and early term infants at the age of 1 year: a follow-up study].

OBJECTIVE: To study the level of neuropsychological development in late preterm infants and early term infants at the age of 1 year. METHODS: A total of 1 257 children with a corrected age of 1 year were enrolled as subjects. According to gestational age at birth, they were divided into an early preterm group (28-33+6 weeks), a late preterm group (34-36+6 weeks), an early term group (37-38+6 weeks), and a full-term group (39-41+6 weeks). Gesell Developmental Schedules were used to assess the neuropsychological development of the children, and the groups were compared in terms of neuropsychological development at the age of 1 year. RESULTS: There were significant differences in the developmental quotients of the five functional areas (adaptability, gross motor, fine motor, language and social ability) between the four groups at the age of 1 year (P<0.05), and the full-term infants had the highest development quotients, followed by the early term infants, the late preterm infants, and the early preterm infants (P<0.05). The full-term infants had the lowest rate of developmental delay in each functional area, while the early preterm infants had the highest rate (P<0.05). Compared with the full-term infants, the early term infants had a higher risk of developmental delay in adaptability (OR=1.796, P<0.05), and the late preterm infants had a higher risk of developmental delay in adaptability (OR=2.651, P<0.05) and fine motor (OR=2.679, P<0.05), while the early preterm infants had a higher risk of developmental delay in adaptability (OR=4.069, P<0.05), fine motor (OR=3.710, P<0.05), and social ability (OR=3.515, P<0.05). CONCLUSIONS: The risk of neuropsychological developmental delay decreases with the increase in gestational age in children at the age of 1 year, with a dose-response effect. There are varying degrees of developmental delay in early term infants and late preterm infants, and health care follow-up for early term infants and late preterm infants should be taken seriously.

High signal-intensity abnormalities in susceptibility-weighted imaging for primary intracerebral hemorrhage.

Purpose: To identify the regularity of signal evolution of intracerebral hemorrhage on susceptibility-weighted imaging (SWI) at different stages compared with T1-weighted imaging (T1WI) and T2-weighted imaging (T2WI). Methods: We retrospectively evaluated a series of 365 patients who underwent T1WI, T2WI, and SWI examination simultaneously or sequentially in our hospital from January 2015 to May 2017. Two neuroradiologists assessed the images and discrepancies between their interpretations were resolved by consensus. Statistical analysis was performed using Chi-squared and Kappa tests. Results: Of the 365 patients on SWI sequence, 94 were enrolled. SWI detected the cases at different stages; T1WI detected 89 cases and T2WI detected 91 cases. The signal intensity of intracerebral hemorrhage on SWI was significantly associated with T1WI imaging and T2WI (χ2 = 4.651; p < 0.05; χ2 = 26.396; p < 0.01, respectively), especially at the late subacute stage. There was moderate consistency between the signal intensity of intracerebral hemorrhage on T2WI and SWI (Kappa coefficient = 0.530). Conclusion: Intracerebral hemorrhage has a varied appearance on SWI, and the evolution of signal of intracerebral hemorrhage on SWI sequence is influenced by T1WI and T2WI. Hematoma detection should be closely combined with clinical manifestation.

Parasubindoles A-G, Seven Eremophilanyl Indoles from the Whole Plant of Parasenecio albus.

Parasubindoles A-G (1-7), seven eremophilanyl indoles with an unprecedented 12 H-cyclopentane[ b]naphthalenespiro-1,3'-indole skeleton, were isolated from the whole plant of Parasenecio albus. Their structures with absolute configurations were elucidated by spectroscopic methods, single-crystal X-ray diffraction, and ECD analyses. Plausible biosynthetic pathways of 1-7 were postulated.

Palladium-catalyzed oxidative C-H/C-H cross-coupling of imidazopyridines with azoles.

An effective palladium-catalyzed oxidative C-H/C-H cross-coupling of imidazopyridines with azoles using air as the oxidant has been developed. This protocol provides a straightforward and operationally simple method for the synthesis of 3-azolyl-imidazopyridines in moderate to good yields and with good functional group tolerance. The biological evaluation revealed that the newly synthesized compounds 3e and 3h exhibit significant in vitro antiproliferative activities against human-derived lung cancer cell lines compared with the positive control, 5-fluorouracil.

Vaccination demonstration zone successfully controls rabies in Guangxi Province, China.

BACKGROUND: Guangxi is the province most seriously affected by rabies virus (RABV) in China. Those most affected by RABV each year are people in rural areas, where dogs are the main cause of human infection with the virus. METHODS: In this study, we established a rabies vaccination demonstration program that included eradication, core, and peripheral areas. This program was implemented for 9 years and comprised three stages: 12 counties in the first stage (2008-2010), 21 counties in the second stage (2011-2013), and then extending to all counties of Guangxi Province in the third stage (2014-2016). The program included a dog vaccination campaign, surveillance of clinically healthy dogs who may be potential RABV carriers, monitoring anti-RABV antibody titers in vaccinated dogs, and compiling and reporting statistics of human rabies cases. RESULTS: The target effectiveness was achieved in the eradication, core, and peripheral areas in all three stages. The vaccination demonstration program successfully promoted RABV vaccination of domestic dogs throughout Guangxi Province by drawing upon the experience gained at key points. Compared with a vaccination coverage rate of 39.42-46.85% in Guangxi Province overall during 2003-2007, this rate gradually increased to 48.98-52.67% in 2008-2010, 60.24-69.67% in 2011-2013, and 70.09-71.53% in 2014-2016, thereby meeting World Health Organization requirements. The total cases of human rabies in the province decreased from 602 in 2004 to 41 cases in 2017. CONCLUSIONS: The present pilot vaccination program obviously increased the rabies vaccination and seroconversion rates, and effectively reduced the spread of rabies from dogs to humans as well as the number of human rabies cases, thus successfully controlling rabies in Guangxi.

Evolutionary analysis of rabies virus isolates from Guangxi Province of southern China.

BACKGROUND: Rabies is a severe epidemic in Guangxi province, China, with hundreds of deaths occurring each year. In the past six decades, rabies has emerged three times in Guangxi, and the province has reported the largest number of rabies cases in China. The domestic dog is the principal vector for rabies, and 95% of human cases are associated with transmission from dogs. RESULTS: To understand the genetic relationship between street rabies virus (RABV) from Guangxi, genetic diversity analysis was performed using RABV isolates collected between 1999 and 2012. The N gene of 42 RABV isolates, and the P and M genes, as well as fragments of the 3' terminus (L1-680) and the polymerase activity module of the L gene (Lpam) of 36 RABV isolates were sequenced. In addition, whole genome sequencing was performed for 5 RABV isolates. There was evidence of topological discrepancy in the phylogenetic trees based on different genes of the RABV isolates. Amino acid variation of the deduced N protein exhibited different patterns to those obtained from the P and M proteins reported here, and the previously reported G protein (Tang H. et al., PLoS Negl Trop Dis, 8(10): e3114, 2014), and L1-680 and Lpam. These RABV isolates were divided into three main branches against fixed strains. CONCLUSION: RABV is prevalent in Guangxi province and strains collected over the last two decades belong mainly to three groups (I, II, III). These RABV isolates reveal genetic diversity. Individual RABV genes from Guangxi exhibit different evolutionary characteristics. The results will have benefits for continuing comprehensive rabies surveillance, prevention and control in China.

MiR-509-3-5p causes aberrant mitosis and anti-proliferative effect by suppression of PLK1 in human lung cancer A549 cells.

MicroRNAs (miRNAs) are potent post-transcriptional regulators of gene expression and play roles in DNA damage response (DDR). PLK1 is identified as a modulator of DNA damage checkpoint. Although down-regulation of PLK1 by certain microRNAs has been reported, little is known about the interplay between PLK1 and miR-509-3-5p in DDR. Here we have demonstrated that miR-509-3-5p repressed PLK1 expression by targeting PLK1 3'-UTR, thereby causing mitotic aberration and growth arrest of human lung cancer A549 cells. Repression of PLK1 by miR-509-3-5p was further evidenced by over-expression of miR-509-3-5p in A549, HepG2 and HCT116p53(-/-) cancer cells, in which PLK1 protein was suppressed. Consistently, miR-509-3-5p was stimulated, while PLK1 protein was down-regulated in A549 cells exposed to CIS and ADR, suggesting that suppression of PLK1 by miR-509-3-5p is a component of CIS/ADR-induced DDR pathway. Flow cytometry and immunofluorescence labeling showed that over-expression of miR-509-3-5p in A549 induced G2/M arrest and aberrant mitosis characterized by abnormal bipolar mitotic spindles, condensed chromosomes, lagging DNA and chromosome bridges. In addition, over-expression of miR-509-3-5p markedly blocked A549 cell proliferation and sensitized the cells to CIS and ADR treatment. Taken together, miR-509-3-5p is a feasible suppressor for cancer by targeting PLK1. Our data may provide aid in potential design of combined chemotherapy and in our better understanding of the roles of microRNAs in response to DNA damage.

Dental caries is negatively correlated with body mass index among 7-9 years old children in Guangzhou, China.

BACKGROUND: Evidence linking caries in primary dentition and children's anthropometric measures is contradictory. We aimed to evaluate the prevalence of primary dental caries and its relationship with body mass index (BMI) among 7-9 years old school children in urban Guangzhou, China. METHODS: This cross-sectional study enrolled 32,461 pupils (14,778 girls and 17,683 boys) aged 7-9 years from 65 elementary schools in Guangzhou. Dental caries was detected according to criteria recommended by the World Health Organization (WHO). The total mean decayed, missing or filled teeth (dmft) of primary dentition were assessed. Weight and height were measured and BMI was calculated. Children were classified into underweight, normal weight, overweight and obesity groups by BMI based on Chinese criteria. Z-score of BMI-for-age (BAZ) was calculated by WHO standardized procedure. Multivariable odds ratios (ORs) and 95 % confidence intervals (CIs) were calculated using logistic regression. Restricted cubic spline regression was applied to evaluate the shape of the relationship between BAZ and primary dental caries. RESULTS: The prevalence of primary dental caries was 30.7 % in total sample. Regarding dmft values, the mean ± standard deviation (SD) in the combined sample were 1.03 ± 2.05 in boys and 0.93 ± 1.92 in girls. Both indices decreased by age. Compared with normal BMI group, children in overweight and obesity groups have 27 % (OR = 0.73, 95 % CI: 0.66-0.81, P < 0.0001) and 34 % (OR = 0.66, 95 % CI: 0.59-0.74, P < 0.0001) lower odds for the presence of primary dental caries after adjustment for age and gender, respectively. Although in general, increased BAZ was associated with decreased risk of dental caries, full-range BAZ was associated with dental caries in an A-shaped manner with a zenith at around -1.4. CONCLUSION: Higher BMI was associated with lower odds of caries; overweight and obese children were more likely to be primary dental caries free among 7-9 years in Guangzhou, China.

[Anisakis simplex larvae: infection status in marine fishes for sale in Shantou].

OBJECTIVE: To investigate the infection status of Anisakis simplex larvae in marine fishes for sale in Shantou. METHODS: Marine fishes were randomly collected from markets in Shantou City from February to December 2013, and then classified. The viscera and muscle of each fish were carefully dissected and thoroughly examined for anisakids. The larvae were examined under a light microscope. The infection rate and intensity of Anisakis simplex larvae were calculated. RESULTS: A total of 382 fish specimens belonging to 52 species were examined. 42 out of 52 species (80.8%) were found infected by A. simplex larvae. The overall infection rate reached 47.4% (181/382), and average 5.5 larvae parasitized per infected fish (995/181). The survival rate of larvae was 100%. The highest infection rate observed was 100% in Scomber australasicus (4/4), Trachurus japonicus (9/9), Decapterus maruadsi (8/8), Lutjanus lutjanus (9/9), Argyrosomus argentatus (4/4), Nibea albiflora (4/4), Nemipterus bathybius (12/12), Trachinocephalus myops (7/7) and Mene maculata (9/9), followed by 16/18 in Pneumatophorus japonicus, 6/7 in Lutjanus ophuysenii and 5/6 in Lutjanus fulvus. A. simplex larvae were not detected in 10 fish species, namely, Megalaspis cordyla, Lutjanus argentimaculatus, Lutjanus fulviflamma, Acanthopagrus australis, Acanthopagrus latus, Plectorhinchus nigrus, Dentex tumifrons, Psenopsis anomala, Scatophagus argus, and Seriola lalandi. The infection intensity was the highest in Lutjanus fulvus (21.0 per fish), followed by Trachinocephalus myops (16.7 per fish), Saurida filamentosa (14.0 per fish) and Mene maculate (10.1 per fish). The lowest infection intensity was found in Rastrelliger kanagurta, Kaiwarinus equula, Atule mate, Lutjanus russellii, Plectorhinchus cinctus, Priacanthus tayenus, Branchiostegus argentatus, Branchiostegus albus, Sphyraena pinguis, Formio niger, Trachinotus blochii, Siganus fuscescens and Choerodon azurio (less than 2 per fish). The highest infection rate (34.3%, 131/382) was found in the mesentery. The infection intensity was highest in pyloric appendage (3.5 per fish). A. simplex larvae were not found in muscle. The highest infection rate (60.2%, 74/123) was found in fishes with body weight of 100-200 g. The infection intensity was highest in fish with body weight of 301-400 g (7.8 per fish). CONCLUSION: The infection rate of A. simplex larvae is high in marine fishes from Shantou markets.

Molecular properties, structure and chiral resolution of secondary metabolites from the leaves of Viburnum chingii.

A chemical investigation of leaves of Viburnum chingii afforded eleven compounds, including one undescribed lignan (1), a pair of known phenylpropanoid enantiomers (2a/2b), and eight known lignans (3-10). Their structures were elucidated by detailed spectroscopic and comparative literature data analysis. The absolute configurations of compounds 1 was determined by comparing the experimental ECD data with the calculated values. The compounds 2a/2b were separated successfully by a chiral chromatographic column. In addition, the acetylcholinesterase (AChE) inhibitory activities of described compounds were evaluated.

Molecular epidemiology of PRRSV from China's Guangxi Province between 2007 and 2009.

Porcine reproductive and respiratory syndrome (PRRS) is considered to be one of the most important infectious diseases affecting livestock. This study used gene sequence analysis of ORF5 and Nsp2 to determine the molecular epidemiology of PRRSV in different parts of the Guangxi province of China. These genes were selected due to their extensive variation within the genome. Out of 189 samples from animals suspected to have PRRS, 145 were PRRSV RNA positive. ORF5 and Nsp2 gene sequence analysis of 31 of these samples showed that all of the Guangxi isolates were of type 2. A phylogenetic tree analysis based on ORF5 showed that the Guangxi isolates were divided into two groups. Most of these were closely related to highly pathogenic strains, showing a 30 amino acid deletion at positions 481 and 533-561 of Nsp2, but an additional unique isolate (GXNN06) possessed a further four amino acid deletion at positions 485-488 of Nsp2.

Two new iridoids and triterpenoid analogues from the leaves of Viburnum chingii and their anti-acetylcholinesterase activity.

Two undescribed split-ring iridoids (1-2) with six known triterpenes (3-8) and one steride (9) were isolated from the Viburnum chingii. Compound 2 possessed an unprecedented split-ring iridoid skeleton formed by electrocyclic reaction and split ring. The structures and absolute configurations of the new iridoids were established by NMR, HRESIMS, and ECD calculations. All the isolated compounds were tested for AChE inhibitory activity. Biologically, 1, 2, 3, 4, and 7 displayed significant AChE effects compared to the positive control donepezil, and have also been subjected to molecular docking studies.

[Determination of 10 carbamate pesticide residues in liquid milk by ultra performance liquid chromatography-tandem mass spectrometry with pass-through solid-phase extraction purification].

Carbamates are used in broad-spectrum insecticides and herbicides, and have highly efficient, low-residue, and long-lasting characteristics. However, this type of pesticide exerts mutagenic, teratogenic, carcinogenic, and other adverse effects, and its frequent use can exceed the recommended scope and limits. Research on the determination of carbamate pesticides mainly focuses on foods of plant origin and pays less attention to foods of animal origin. The methods for carbamate determination described in the current national standards have complicated operating procedures and low efficiency. Therefore, highly efficient and accurate methods for carbamate detection in milk must be established. In this work, a rapid method based on pass-through solid-phase extraction (SPE) purification coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous determination of 10 carbamate pesticides in liquid milk. The pretreatment and instrument methods were systematically optimized. The milk sample was extracted with acetonitrile, and then purified using a Captiva EMR-Lipid filtration kit. The purified extract was separated on an ACQUITY UPLC BEH C18 column with mobile phase of methanol and 0.1% formic acid aqueous solution in gradient elution. The flow rate was 0.3 mL/min. Column temperature was 35 ℃. Quantitative analysis was performed using the external standard method with matrix matching curves. The 10 carbamate pesticides showed good linear relationships in the mass concentration range of 2-200 µg/L, with correlation coefficients greater than 0.999. The limits of detection (LODs) and quantification (LOQs) for the 10 carbamate pesticides were 0.045-0.23 and 0.15-0.77 µg/kg, respectively. Recovery tests were conducted using the blank-matrix method at three spiked levels of 15, 50, and 100 µg/kg, and good recoveries for the 10 carbamate pesticides were obtained. In particular, the recoveries for the three spiked levels of 15, 50, and 100 µg/kg were 68.7%-93.3% with relative standard deviations (RSDs) of 1.8%-8.0%. The proposed method is efficient, convenient, accurate, and suitable for the rapid detection of the 10 carbamate pesticides in liquid milk. Compared with the conventional NH2 and ENVITM-18 SPE columns used in the national standard determination method, the proposed method demonstrated better purification effects. The recoveries for aldicarb sulfoxide, aldicarb sulfone, methomyl, and carbaryl after purification using the Captiva EMR-Lipid kit increased from 60% to 80%. Thus, the proposed method is suitable for targets with strong polarity and gives measurement results with good repeatability and accuracy.

Effectiveness of group acceptance and commitment therapy in treating depression for acute stroke patients.

OBJECTIVES: To date, the effectiveness of acceptance and commitment therapy (ACT) for acute stroke patients has not been well recognized. The study aimed to discover the effectiveness of group-based ACT in treating depression for acute stroke patients. METHODS: We conducted a randomized controlled trial with 140 acute stroke patients with depression. The ACT intervention comprised seven sessions, of 45-60 min over 4 weeks. Data were collected pre- and post-intervention and at 3-month follow-up, assessing depression, health-related quality of life (HRQoL), psychological flexibility, cognitive fusion, sleep quality, and confidence. RESULTS: Overall, 99.3% of the included patients were assessed as having mild depression. The ACT intervention significantly reduced depression in acute stroke patients in comparison with the control group post-intervention and at 3 months (partial η 2 = . 306 $\eta^{2}=.306$ ). Additionally ACT significantly improved HRQoL-mental component summary, sleep quality, psychological flexibility, cognitive fusion, and confidence compared with control group. CONCLUSIONS: ACT is effective in treating acute stroke patients with depression, and the efficacy was maintained at 3-month follow-up.

In vitro infection with classical swine fever virus inhibits the transcription of immune response genes.

BACKGROUND: Classical swine fever virus (CSFV) can evade the immune response and establish chronic infection under natural and experimental conditions. Some genes related to antigen processing and presentation and to cytokine regulation are known to be involved in this response, but the precise mechanism through which each gene responds to CSFV infection remains unclear. RESULTS: In this study, the amplification standard curve and corresponding linear regression equations for the genes SLA-2, TAP1, SLA-DR, Ii, CD40, CD80, CD86, IFN-α, and IFN-ß were established successfully. Real-time RT-PCR was used to quantify the immune response gene transcription in PK-15 cells post CSFV infection. Results showed that: (1) immune response genes were generally down-regulated as a result of CSFV infection, and (2) the expression of SLA-2, SLA-DR, Ii and CD80 was significantly decreased (p < 0.001). CONCLUSION: We conclude that in vitro infection with CSFV inhibits the transcription of host immune response genes. These findings may facilitate the development of effective strategies for controlling CSF.

[Progress of Non-Factor Products in Hemophilia Treatment--Review].

Traditional replacement therapy is the main treatment method of hemophilia, while inhibitor generation makes replacement therapy ineffective. The emergence of non-factor therapy brings new hope for the treatment of patients with inhibitor. Non-factor products mainly achieve therapeutic purpose by imitating the function of coagulation factor Ⅷ, inhibiting the function of anti-tissue factor pathway inhibitors, the expression of antithrombin mRNA, and the function of activated protein C. This paper reviews the latest research progress of non-factor products in the treatment of hemophilia.