Abstract To study the residue behavior of pyraclostrobin in wax gourd, and evaluate the risk of dietary intake. Wax gourd samples were extracted with acetonitrile, purified with PSA and C18, detected by HPLC-MS/MS. In 2017, the supervised field trials of pyraclostrobin in wax gourd were carried out in Henan, Jiangsu, Sichuan, Beijing, Hunan, and Guangxi. The STMR of pyraclostrobin in wax gourd was acquired, and then the risk of dietary intake was evaluated. At the three spiked levels of 0.01, 0.1 and 1.0 mg kg-1, the average recoveries ranged from 95% to 104%, the RSDs ranged from 2.1% to 3.8%, and LOQ was 0.01 mg kg-1. The dissipation of pyraclostrobin in wax gourd in Sichuan fitted to the first order kinetics with the half-life of 4.1d. The 250 gL-1 emulsifiable concentrate of pyraclostrobin was sprayed at 150 and 225 g a.i./hm2 for 2-3 times on wax gourd. The final residue level of pyraclostrobin in wax gourd was between <LOQ and 0.11 mgkg-1. The national estimated daily intake (NEDI) of pyraclostrobin was 0.3722 mg, accounting for 19.69% of the acceptable daily intake (ADI) for general population, which means that it will not pose unacceptable risks to the health of the general population.

Resumo Fundamento A relação direta entre a doença arterial coronariana (DAC) e o câncer de pulmão não é bem conhecida. Objetivo Investigar a associação entre a gravidade anatômica da DAC e do câncer de pulmão. Métodos Trezentos pacientes, incluindo 75 recém-diagnosticados com câncer de pulmão e 225 pacientes correspondentes sem câncer, foram submetidos à angiografia coronária durante a internação, sem intervenção coronária percutânea (ICP) prévia nem enxerto de bypass da artéria coronária (CABG). O escore SYNTAX foi utilizado para avaliar a gravidade da DAC. Uma pontuação alta no escore foi definida como > 15 (o maior quartil do escore SYNTAX). O teste de tendência de Cochran-Armitage foi utilizado para verificar a distribuição dos escores dos pacientes. Uma análise de regressão logística foi utilizada para avaliar a associação entre a gravidade da DAC e o câncer de pulmão. Os valores de p foram estabelecidos quando o nível de significância era 5%. Resultados A tendência de distribuição dos escores SYNTAX dos pacientes por quartis foi diferente entre aqueles com câncer de pulmão e controles (do quartil mais baixo ao mais alto: 20,0%; 20,0%; 24,0%; 36,0% vs. 26,7%; 26,2%; 25,8%; 21,3%; p=0,022). A pontuação no escore SYNTAX foi mais alta em pacientes com câncer do que nos pacientes controle (36,0% vs. 21,3%, p=0,011).O maior quartil do escore demonstrou mais riscos de desenvolver câncer de pulmão em comparação ao quartil mais baixo (OR: 2.250, IC95%: 1.077 a 4.699 ; P -trend= 0,016). Após ajustes, os pacientes no maior quartil do escore SYNTAX tinham mais risco de desenvolver câncer de pulmão (OR: 2.1o49, IC95%: 1.008 a 4.584; P -trend= 0,028). Pacientes com escores SYNTAX alto (> 15) tinham 1.985 mais chances de ter câncer de pulmão (IC95%: 1.105–3.563, P= 0,022). Conclusão A gravidade anatômica da DAC está associada ao risco de câncer de pulmão, o que indica que um rastreamento completo deste tipo de câncer possa ser mais significativo entre pacientes com DAC.

Abstract Background The direct relationship between coronary artery disease (CAD) and lung cancer is not well known. Objective To investigate the association between the anatomical severity of CAD and lung cancer. Methods Three-hundred study patients, including 75 recently diagnosed lung cancer patients and 225 matched non-cancer patients, underwent coronary angiography during hospitalization without previous percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). The SYNTAX score (SXscore) was used to assess the severity of CAD. A high SXscore (SXhigh) grade was defined as SXscore > 15 (the highest quartile of the SXscore). The Cochran-Armitage test for trend was used to assess the distribution of patients’ SXscores. Logistic regression analysis was used to assess the association between the severity of CAD and lung cancer. P-values were set when significance level was 5%. Results The distribution trend of patients’ SXscore by quartiles was different between lung cancer patients and control patients (from the lowest to the highest quartile: 20.0%, 20.0%, 24.0%, 36.0% vs. 26.7%, 26.2%, 25.8%, 21.3%, p=0.022). The SX high rate was higher in lung cancer patients than in control patients (36.0% vs. 21.3%, p=0.011).The highest quartile of the SXscore showed higher risk of lung cancer in comparison to the lowest quartile (OR: 2,250, 95%CI: 1,077 to 4,699 ; P-trend= 0.016). After adjustment, patients in the highest quartile of the SXscore had higher risk of lung cancer (OR: 2,149, 95%CI: 1,008 to 4,584; P-trend= 0.028). Patients with high SXscore (> 15) had 1,985 times more chances of having lung cancer (95%CI: 1,105–3,563, P= 0.022). Conclusions The anatomical severity of CAD is associated with the risk of lung cancer, which indicates that a thorough lung cancer screening may be significant among severe CAD patients.

ABSTRACT A 35-year-old immunocompetent woman from southern China went to the hand surgery clinic with a six-month history of progressive swelling in her right index finger. She had been pinched by a lobster and had received several treatments without any improvement. Pus specimens were taken from the swollen parts of her finger, and the pathology showed granulomatous inflammation. Ziehl–Neelsen staining revealed positive bacillus in the pus specimens. The bacteria grew well on Columbia blood agar. However, the MALDI-TOF MS and 16S rRNA gene sequencing were not able to distinguish between Mycobacterium marinum and Mycobacterium ulcerans because of their close genetic relationship. Photochromogenicity testing can help differentiate between these species based on the alteration in colony color after light exposure. For our patient, the colonies turned yellow after 18h of incubation in the sun, identifying the species as M. marinum. Besides surgical drainage, the patient received rifampicin and clarithromycin for three months, and her symptoms resolved without relapse after six months of follow-up.

Abstract Background: To assess the expression of T-box transcription factor 4 (TBX4) during the anorectal development in normal and ethylenethiourea (ETU)-induced anorectal malformations (ARM) rat embryos. Methods: Anorectal malformations was induced by ETU on the 10th gestational day (E10) in rat embryos. Spatiotemporal expression of TBX4 was evaluated in normal (n = 490) and ETU-induced ARM rat embryos (n = 455) from E13 to E16 by immunohistochemical staining, Western blot analysis and real-time RT-PCR. Results: In the normal embryos, immunohistochemical staining revealed that TBX4 expression was detected in the epithelium of hindgut and urorectal septum (URS) on E13. TBX4-immunopositive cells were increased significantly in the epithelium of hindgut and URS, the future anal orifice part of cloacal membrane on E14. On E15, abundant stained cells were observed in the rectum, URS and dorsal cloacal membrane and the expression of positive cells reached its peak. On E16, only sporadic positive cells were distributed in the epithelium of the distal rectum. In the ARM embryos, the hindgut/rectum, URS and dorsal cloacal membrane were faint for TBX4 immunohistochemical staining. In the normal group, TBX4 protein and mRNA expression showed time-dependent changes in the hindgut/rectum from E13 to E16 on Western blot and real-time RT-PCR. On E13 and E15, the expression level of TBX4 mRNA in the ARM group was significantly lower than that in the normal group (P < 0.05). On E15, the expression level of TBX4 protein in the ARM group was significantly lower than that in the normal group (P < 0.05). Conclusions: The expression of TBX4 was downregulated in ETU-induced ARM embryos, which may play important roles in the pathogenesis of anorectal development.

ABSTRACT The carapace of the tortoise Chinemys reevesii is an ingredient of “Guijia”, a traditional Chinese medicine. However, C. reevesii is difficult to raise in aquaculture and is rare in the wild. Counterfeit tablets are made from carapaces of other species. In addition to C. reevesii, other species including Mauremys sinensis, Indotestudo elongate and Trachemys scripta have been used in Plastrum Testudinis as well. After processing, these carapaces are difficult to identify on the basis of morphological characteristics, which impedes law enforcement. Our study used DNA barcoding technology to identify C. reevesii and its substitutes. We extracted concentrated genomic DNA for PCR amplification. Based on the analysis of 61 full-length COI sequences, we designed four pairs of mini-barcode primers: Tu-A, Tu-B, Tu-C and Tu-D. The Tu-B primers sequenced genomic DNA with a success rate of 76.47%, and the Tu-D primers sequenced genomic DNA with a success rate of 88.24%. The identification efficiency of these two mini-barcodes was 70.59% and 64.71%, and the overall identification efficiency was approximately 76.47%. Similarly, a set of mini barcode systems was generated, which may provide an effective and low-cost method for the identification of authentic tortoise shells.

Abstract Wide compatibility and erect panicles are important traits for heterosis and ideal plant type breeding in rice and accordingly are targets for rice yield improvement. In this study, a multiplex PCR system based on functional markers was developed to simultaneously identify genotypes of the wide compatibility allele S5-n and the erect panicle allele dep1. Forty-nine rice varieties in the Huang-Huai-Hai region of China were identified using this system. Thirty-three varieties contained the erect panicle allele dep1 and two varieties contained the wide compatibility allele S5-n. The frequency of dep1 was obviously higher than that of S5-n. The multiplex PCR method was also tested using an F2 segregating population and was found to be simple, efficient, and reliable. It will be useful in marker-assisted selection based on the wide compatibility allele S5-n and erect panicle allele dep1 to breed super rice varieties using indica and japonica heterosis.

The endangered causes of Taxus chinensis var. mairei in the Taihang Mountains are analyzed in three sides in connection with the situation that is resources increasing attenuation. The first is biological factors such as pollination barriers, deeply dormancy seed, cannot vegetative propagation under natural conditions, poor adaptability of seedling to environment and slow growth. The second is environmental factors such as very limited distribution environment and position in community. The third is interference of persons and other animals. According to these factors, we provide three measures to protect Taxus chinensis var. mairei in three sides that protect existing resources, breed subsequent resources and find new pathway of producing taxol.

Las razones por las que la especie Taxus chinensis var. mairei está en peligro de extinción en las montañas de Taihang giran en torno a tres ejes distintos en relación a la disminución de ejemplares. En primer lugar contamos con factores biológicos, como las barreras de polinización, el alto grado de inactividad de sus semillas, la imposibilidad de propagación vegetal por medios naturales, la escasa adaptación de los vástagos al medio y su lento crecimiento. El segundo factor es de carácter ambiental: medio de distribución y posición en la comunidad muy limitados. Por último, la tercera causa es la interferencia de personas y animales. De acuerdo con estos factores, proponemos tres medidas para proteger la especie Taxus chinensis var. mairei con el fin de conservar los ejemplares existentes, cultivar nuevos ejemplares y encontrar nuevas vías para producir taxol.

Background: Wheat 1BL/1RS translocation lines are planted around the world for their disease resistance and high yield. Most of them are poor in bread making, which is partially caused by ω-secalins that are encoded by the ω-secalin gene family, which is located on the short arm of rye chromosome 1R (1RS). However, information on the structure and evolution of the ω-secalin gene family is still limited. Results: We first generated a physical map of the ω-secalin gene family covering 195 kb of the Sec-1 locus based on sequencing three bacterial artificial chromosome (BAC) clones of the 1BL/1RS translocation wheat cultivar Shimai 15. A BAC contig was constructed spanning 168 kb of the Sec-1 locus on 1RS. Twelve ω-secalin genes were arranged in a head-to-tail fashion, separated by 8.2-21.6 kb spacers on the contig, whereas six other ω-secalin genes were arranged head-to-tail, separated by 8.2-8.4 kb of spacers on clone BAC125. The 18 ω-secalin genes can be classified into six types among which eight ω-secalin genes were expressed during seed development. The ω-secalin genes with the 1074-bp open reading frame (ORF) represented the main population. Except for two pseudogenes, the N-terminal of the ω-secalin gene was conserved, whereas variations in the C-terminal led to a change in ORF length. The spacers can be sorted into two classes. Class-1 spacers contained conserved and non-conservative sequences. Conclusion: The ω-secalin gene family consisted of at least 18 members in the 1BL/1RS translocation line cv. Shimai 15. Eight ω-secalin genes were expressed during seed development. Eighteen members may originate from a progenitor with a 1,074-bp ORF. The spacers differed in length and sequence conservation.

The objective of this study was to isolate endophytic fungi producing paclitaxel from yew for the purpose of paclitaxel manufacture. Surface sterilized bark of Taxus wallichiana var. mairei was used as source material and potato dextrose agar culture medium was used in isolation of endophytic fungi. Fungal cultures were extracted with a mixture of chloroform / methanol (1:1, v/v) and the paclitaxel in the extracts was determined and authenticated with LC-MS. An endophytic fungus that produced paclitaxel was identified by ITS rDNA and 26S D1/D2 rDNA sequencing. The results showed that a total of 435 endophytic fungal strains were isolated from T. wallichiana var. mairei and purified. Only one of these strains produced paclitaxel and it belongs to Fusarium. The paclitaxel productivity in whole PDB culture and that in spent culture medium from this strain is 0.0153 mg/L and 0.0119 mg/L respectively. The paclitaxel content in dry mycelium is 0.27 mg/kg. This isolated endophytic fungus produced paclitaxel at a considerable level and shows potentiality as a producing strain for paclitaxel manufacture after strain improvement.

El objetivo de este estudio fue aislar hongos endofíticos productores de paclitaxel a partir de tejo con el propósito de fabricar paclitaxel. Se utilizó la superficie de la corteza esterilizada de Taxus wallichiana var. mairei como material de origen y dextrosa de patata en medio de cultivo de agar para el aislamiento de hongos endófitos. Los cultivos de hongos se extrajeron con una mezcla de cloroformo / metanol (1:1, v/v) y el paclitaxel en los extractos se determinó y autentificó con LC-MS. Un hongo endófito que produjo paclitaxel fue identificado por su ADNr 26S y secuenciación D1/D2 ADNr. Los resultados mostraron que un total de 435 cepas de hongos endófitos se aislaron y purificarón a partir de T. wallichiana var. mairei. Solo una de estas cepas produce paclitaxel y pertenece a Fusarium. La productividad del cultivo de paclitaxel procedente de esta cepa es 0,0153 mg/L y 0,0119 mg/L, respectivamente. El contenido de paclitaxel en micelio seco es 0,27 mg/kg. Este aislado de hongos endófitos produjo paclitaxel a un nivel considerable y muestra potencial como cepa para la fabricación de paclitaxel después de llevar a cabo una mejora de las cepas.

Este trabalho objetiva o estudo da patogênese de deficiência no aprendizado e memória de prole de ratos resultante da anestesia maternal por enflurano, por meio da expressão da subunidade 2B do receptor do ácidoN-metil-D-aspártico (NR2B) no hipocampo dos filhotes. Dividiram-se, aleatoriamente, 30 fêmeas de ratos Sprague-Dawley em três grupos: controle (grupo C), exposição ao enflurano por 4 h (grupo E1) e por 8 h (grupo E2). De oito a 10 dias após o início da gravidez, os ratos dos grupos E1 e E2 inalaram enflurano 1,7% em 2 L/min de oxigênio, por 4 h e 8 h, respectivamente. Ratos do grupo C inalaram apenas 2 L/min de oxigênio. O labirinto de água de Morris foi empregado para analisar as funções de aprendizado e memória da cria em 20 e 30 dias após o nascimento. Utilizaram-se ensaios de RT-PCR e de imuno-histoquímica para medir os níveis de mRNA e expressão da proteína do NR2B, respectivamente. Em comparação com os ratos controle do grupo C, aqueles dos grupos E1 e E2 exibiram latências de escape mais longas, menor número de travessias na plataforma e menos tempo gasto no quadrante alvo no teste de exploração espacial (P < 0,05). Adicionalmente, os níveis de expressão de mRNA e de proteína do NR2B no hipocampo dos filhotes nos grupos E1 e E2 estavam reduzidos (P < 0,05). Não se observaram diferenças significativas entre os grupos E1 e E2 (P < 0,05) quanto aos níveis de mRNA e à expressão de proteína de NR2B. A função cognitiva dos filhotes é prejudicada quando as mães são expostas ao enflurano durante o início da gravidez. O mecanismo possível para esse efeito está relacionado à diminuição na expressão de NR2B.

This work aims to study the pathogenesis of learning and memory impairment in offspring rats resulting from maternal enflurane anesthesia by focusing on the expression of the N-methyl-d-aspartic acid receptor subunit 2B (NR2B) in the hippocampus of the offspring. Thirty female Sprague-Dawley rats were randomly divided into three groups: control (C group), 4 h enflurane exposure (E1 group), and 8 h enflurane exposure (E2 group) groups. Eight to ten days after the initiation of pregnancy, rats from the E1 and E2 groups were allowed to inhale 1.7% enflurane in 2 L/min oxygen for 4 h and 8 h, respectively. Rats from the C group were allowed to inhale 2 L/min of oxygen only. The Morris water maze was used to assay the learning and memory function of the offspring on postnatal days 20 and 30. RT-PCR and immunohistochemistry assays were then used to measure the mRNA levels and protein expression of NR2B, respectively. Relative to offspring rats from the C group, those from the E1 and E2 groups exhibited longer escape latencies, lesser number of crossings over the platform, and less time spent in the target quadrant in the spatial exploration test (P < 0.05). In addition, the mRNA and protein expression levels of NR2B in the hippocampus of offspring rats in the E1 and E2 groups were down-regulated (P < 0.05). No significant differences between the E1 and E2 groups were observed (P > 0.05) in terms of mRNA levels and protein expression of NR2B. The cognitive function of the offspring is impaired when maternal rats are exposed to enflurane during early pregnancy. A possible mechanism of this effect is related to the down-regulation of NR2B expression.

Objective: The purpose of this paper is to comprehensively evaluate the quality of Chishao. Methods: In the experiment of this paper, the fingerprint spectrums of Chishao in all locations are established by RP-HPLC and the model of principle component analysis with the RP-HPLC peak areas is established. Results: The quality of Chishao in the northern part of China or that made of Paeonia lactiflora is better than that of these in others or that made of other species. The quality of Chishao comes from P. veitchii is in the middle class and is better than those that comes from P. obovata, P. mairei and P. anomala. The results are consistent with traditional views of the quality of this plant. These results indicates that principal component analysis (PCA) can be used as an effective and economic method to evaluate the quality of Chishao, and may be extended to other Chinese medicinal plants. Conclusions: Due to the complex basis of the efficacy of Traditional Chinese Medicine (TCM), the method such as PCA of several chemical components appears to be a more appropriate method for the quality evaluation of TCM in contrast to the determination of a single or few chemicals.

Objetivo: El propósito de este artículo es la evaluación cualitativa extensa de Chishao. Métodos: En el experimento de este trabajo, se establecen los espectros de identificación de Chishao en todas las localizaciones mediante RP-HPLC y el método del análisis de componentes principales con las áreas pico de RP-HPLC. Resultados: La calidad de Chishao en el norte de China o del procedente de Paeonia lactiflora es mejor que la de otras localizaciones o procedente de P. obovata, P. mairei y P. anomala. Los resultados son congruentes con la impresión tradicional de la calidad de esta planta. Estos resultados indican que el análisis de los componentes principales (ACP) puede utilizarse como método eficaz y económico para evaluar la calidad de Chishao y podría aplicarse a otras plantas medicinales chinas. Conclusiones: Dada la complicada base de la eficacia de la Medicina tradicional china (MTC), un método como el ACP para diversos componentes químicos parece ser más adecuado para la evaluación de la calidad de la MTC en comparación con la determinación de un único o unos pocos agentes químicos.

Cryptococcus spp. are common causes of mycoses in immunocompromised patients. To determine the drug susceptibilities of clinical Cryptococcus spp. isolates, the characteristics of 61 clinical Cryptococcus spp. complex isolates and their antifungal susceptibilities were investigated, including 52 C. neoformans and 9 C. gattii isolates collected at Shanghai between 1993 and 2009. Antifungal susceptibility of clinical isolates to amphotericin B, fluconazole, itraconazole, and flucytosine were determined by the microdilution method M27-A2 and the ATB FUNGUS 3 kit. The 90% minimum inhibitory concentration (MIC90) and susceptibility ranges were as follows: 1 (0.0625-1) µg/mL for amphotericin B, 4 (0.125-16) µg/ mL for fluconazole, 0.25 (0.0313-4) µg/mL for itraconazole, and 4 (0.125-8) µg/mL for flucytosine. Fluconazole, itraconazole, and flucytosine have excellent in vitro activity against all tested clinical Cryptococcus spp., and we also found a high rate of tolerance to amphotericin B (MICs ranging from 0.55-1 µg/mL). Furthermore, C. neoformans isolates from acquired immune deficiency syndrome (AIDS) patients were less susceptible to fluconazole and flucytosine than those from non-AIDS patients. These data suggest that use of amphotericin B may lead to tolerance or resistance of the pathogen over time. There were also no significant associations between species, genotypes, and in vitro susceptibilities of these clinical isolates.

Making use of the gene resources of wild type peanuts is a way to increase the genetic diversity of the cultivars. Marker assisted selection (MAS) could shorten the process of inter-specific hybridization and provide a possible way to remove the undesirable traits. However, the limited number of molecular markers available in peanut retarded its MAS process. We started a peanut ESTs (Expressed Sequence Tags) project aiming at cloning genes with agronomic importance and developing molecular markers. In this study we found 610 ESTs that contained one or more SSRs from 12,000 peanut ESTs. The most abundant SSRs in peanut are trinucleotides (66.3%) SSRs and followed by dinucleotide (28.8%) SSRs. AG/TC (10.7%) repeat was the most abundant and followed by CT/GA (9.0%), CTT/GAA (7.4%), and AAG/TTC (7.3%) repeats. Ninety-four SSR containing ESTs were randomly selected for primer design and synthesis, of which 33 pairs could generate good amplification and were used for polymorphism assessment. Results showed that polymorphism was very low in cultivars, while high level of polymorphism was revealed in wild type peanuts.