Substantial Epigenetic Variation Causing Flower Color Chimerism in the Ornamental Tree Prunus mume Revealed by Single Base Resolution Methylome Detection and Transcriptome Sequencing.

Epigenetic changes caused by methylcytosine modification participate in gene regulation and transposable element (TE) repression, resulting in phenotypic variation. Although the effects of DNA methylation and TE repression on flower, fruit, seed coat, and leaf pigmentation have been investigated, little is known about the relationship between methylation and flower color chimerism. In this study, we used a comparative methylomic⁻transcriptomic approach to explore the molecular mechanism responsible for chimeric flowers in Prunus mume "Danban Tiaozhi". High-performance liquid chromatography-electrospray ionization mass spectrometry revealed that the variation in white (WT) and red (RT) petal tissues in this species is directly due to the accumulation of anthocyanins, i.e., cyanidin 3,5-O-diglucoside, cyanidin 3-O-glucoside, and peonidin 3-O-glucoside. We next mapped the first-ever generated methylomes of P. mume, and found that 11.29⁻14.83% of the genomic cytosine sites were methylated. We also determined that gene expression was negatively correlated with methylcytosine level in general, and uncovered significant epigenetic variation between WT and RT. Furthermore, we detected differentially methylated regions (DMRs) and DMR-related genes between WT and RT, and concluded that many of these genes, including differentially expressed genes (DEGs) and transcription factor genes, are critical participants in the anthocyanin regulatory pathway. Importantly, some of the associated DEGs harbored TE insertions that were also modified by methylcytosine. The above evidence suggest that flower color chimerism in P. mume is induced by the DNA methylation of critical genes and TEs.

[Risk Assessment of Snail Output Via the Mud Balls of Transplanted Seedlings in a Nursery Stock Park in Middle Region of Zhejiang Province].

Objective: To investigate the distribution of snails in a nursery stock park in the middle region of Zhejiang Province and assess the risk of snail output via the mud balls of transplanted seedlings, to provide scientific data for making strategies for snail control. Methods: We selected three species of seedlings including Osmanthus fragrans (a large tree), Camellia sasanqua (a small tree), and Purpus privet (a type of shrub) in a nursery stock park in a snail-positive middle region of Zhejiang Province during 2014-2016 to calculate the areas of regions with snails and the density of living snails. In 30 trees of each species, the distribution of snails within the seedlings ground diameter (radius of investigation, 100 cm for Osmanthus fragrans; 30 cm for Camellia Sasanqua and Purpus Privet) and in different soil layers (surface and superficial layers, 0-3 cm; deep layer, 3-10 cm) was assessed. In addition, the presence of snails in mud balls of 50 trees of Photinia fraseri (a small tree with high density of snails) was investigated to assess the risk of snail output. Results: In the planting areas of Osmanthus fragrans（3 930 m2）, Camellia sasanqua（2 000 m2）, and Purpus privet （1 700 m2）, the areas of snail-positive regions were 200, 900 and 800 m2, respectively, with the density of living snails being 0.08, 0.56 and 0.55/0.1 m2. For Osmanthus fragrans, Camellia sasanqua and Purpus privet, 238, 654 and 645 snails were detected respectively within their seedlings ground diameter, including 159（66.8%）, 461（70.5%） and 376 （58.3%） snails in the surface layer, respectively, which were significantly higher than those in the superficial and deep layers（P＜0.01）. Snails were found in all the 50 trees of Photinia fraseri（3 726 snails, 706 adult snails and 3 020 immature snails, 75 snails/tree on average）. Conclusion: There is a high density of snails in the nursery stock park in the middle region of Zhejiang Province. The snails are distributed mainly in the surface layer, suggesting a risk of snail output through mud balls.

[Interpretation of Criteria for Diagnosis of Toxoplasmosis].

The criteria for Diagnosis of Toxoplasmosis (WS/T 486-2015)（referred to as the Criteria） was compiled following the Management Measures for Health Criteria and GBT 1.1-2009 Standardization Working Guidelines. The Criteria is composed of six chapters, including the range of application, terms and definitions, diagnostic principle, diagnostic standard, and differential diagnosis. Four informative appendices (etiology, epidemiology, clinical manifestation, and differential diagnosis) and one normative appendix (laboratory examination) are appended. The Criteria was issued by the National Health and Family Planning Commission of the People's Republic of China through No.21 of Chinese Health Announcement in 2015. The Criteria provides for the first time technical reference for diagnosis of toxoplasmosis in medical institutions and disease control institutions.

[Evaluation on molluscicidal effect of chlorosalicylicamide on Oncomelania hupensis in schistosomiaisis endemic areas of eight provinces in China].

OBJECTIVE: To evaluate the effects of a novel molluscicide, the salt quinoid-2', 5-dichloro-4'-nitrosalicylanilide from niclosamide (LDS), with 10% wettable powder, in main schistosomiasis epidemic areas of China, including Hunan, Jiangxi, Hubei, Anhui, Jiangsu, Sichuan, Yunnan and Zhejiang Province. METHODS: In the immersion test, 6 effective concentrations of 10% LDS were tested respectively: 0.1, 0.2, 0.4, 0.6, 0.8, 1.0 g/m3 in the field; at the same time, 50% wettable powder of niclosamide ethanolamine salt (WPN) with effective concentrations of 1.0 g/m was used as the molluscicide control, and the fresh water as the blank control, then the mortality rates of 0. hupensis snails were recorded at 24 h, 48 h and 72 h after the immersion. In the spraying test and powder-spraying test, 5 effective dosages of 10% LDS were tested respectively: 0.2, 0.4, 0.6, 0.8, 1.0 g/m2, while 50% WPN 1.0 g/m2 was used as the molluscicide control, and the fresh water as the blank control in the field for 1 d, 3 d and 7 d, then the mortality rates of O. hupensis snails were recorded at 1 d, 3 d and 7 d after the spraying and powder-spraying. RESULTS: The snail mortality rates of LDS using the immersion test for 72 h were more than 95% in the field of eight provinces (0.1 g/m in Sichuan, Jiangxi, Jiangsu and Zhejiang provinces, 0.2 g/m3 in Yunnan, Hunan and Hubei provinces, and 0.4 g/min Anhui Province); the snail mortality rates of LDS using the spraying test for 7 d were more than 85% (0.2 g/m2 in Hunan, Hubei, Jiangxi and Zhejiang provinces, 0.4 g/m2 in Sichuan and Anhui provinces, 0.6 g/m2 in Yunnan and Jiangsu provinces). The snail mortality rates of LDS the powder-spraying test for 7 d were more than 85% (0.6 g/m2 in Yunnan, Sichuan, Hubei, Jiangxi, Anhui, Jiangsu and Zhejiang provinces). According to the standards of "Efficacy test methods and evaluation of molluscicide for pesticide registration (NY/T 1617-2008)", LDS is a qualified molluscicide. CONCLUSIONS: LDS has good molluscicidal effects through the immersion, spraying and powder-spraying test in the fields. It is suitable for a variety of environments to control O. hupensis snails of schistosomiasis endemic areas in China. The recommended dosages of LDS are 0.1-0.2 g/m3 by the immersion method, 0.2-0.4 g/m2 by the spraying method, and 0.4-0.6 g/m2 by the powder-spraying method in the fields.

[Strategy of solidification and surveillance for schistosomiasis in transmission-interrupted provinces in China].

In schistosomiasis transmission-interrupted provinces in China, the endogenous source of infection has been basically cleared, and the exogenous source of infection is considered as a major threat. The residual snail area has been increasing in different degrees, and imported snails are regarded as a potential risk. So the possibility of schistosomiasis recrudescence still remains. According to the endemic characteristics, the integrated solidification strategy with emphasis on control of residual snails and imported cases should be taken in the future.

[Quality control assessments of feces examination for schistosomiasis in province-level laboratories of Zhejiang Province].

OBJECTIVE: To understand and evaluate the quality of feces examination for schistosomiasis in province-level laboratories of Zhejiang Province. METHOD: With the single-blind method, the stool samples were detected by the stool hatching method and sediment detection method. RESULTS: In the 3 quality control assessments in 2006, 2008 and 2009, most laboratories finished the examinations on time. The accordance rates of detections were 88.9%, 100% and 93.9%, respectively. CONCLUSION: The province-level laboratories for schistosomiasis feces examination of Zhejiang Province is coming into standardization, and the techniques of schistosomiasis feces examination are optimized gradually.

Development of a rapid dipstick with latex immunochromatographic assay (DLIA) for diagnosis of schistosomiasis japonica.

BACKGROUND: Schistosomiasis japonica (schistosomiasis) is a zoonosis that can seriously affect human health. At present, the immunodiagnostic assays for schistosomiasis detection are time-consuming and require well-trained personnel and special instruments, which can limit their use in the field. Thus, there is a pressing need for a simple and rapid immunoassay to screen patients on a large scale. In this study, we developed a novel rapid dipstick with latex immunochromatographic assay (DLIA) to detect anti-Schisaosoma japonicum antibodies in human serum. RESULTS: Using latex microspheres as a color probe, DLIA was established to test standard positive and negative sera, in comparison with the classical enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of DLIA were 95.10% (97/102) and 94.91% (261/275), respectively. The cross-reaction rates with clonorchiosis, intestinal nematodes, Angiostrongylus cantonensis and paragonimiasis were 0, 0, 0 and 42.11% respectively. All the results showed no significant difference to the ELISA. In field tests, 333 human serum samples from an endemic area were tested with DLIA, and compared with ELISA and Kato-Katz method. There was no significant difference between DLIA and ELISA on positive and negative rates of detection; however, significant differences existed between DLIA and Kato-Katz method, and between ELISA and Kato-Katz method. The kappa value between DLIA and ELISA was 0.90. CONCLUSIONS: This is the first study in which DLIA was used to detect anti-Schistosoma japonicum antibody. The results show that DLIA is a simple, rapid, convenient, sensitive and specific assay for the diagnosis of schistosomiasis and is therefore very suitable for large-scale field applications and clinical detection.

[Recent investigation and treatment of 1 060 advanced schistosomiasis cases in Zhejiang Province].

OBJECTIVE: To understand the current status of advanced schistosomiasis patients, deliver medical treatment, and improve the case management in Zhejiang Province. METHODS: The cases previously diagnosed or suspected as advanced schistosomiasis in the province were the subjects of the investigation. Questionnairing (demographical information, disease history, etc.), clinical examination (ascites syndrome, abdominal palpation), laboratory examination (blood biochemistry, anti-Schistosoma japonicum antibody, eggs), and ultrasonography of the abdomen were used confirming the diagnosis. Treatment was given to the patients. RESULTS: There were 1 060 advanced schistosomiasis patients in 32 counties of 7 prefectures in the province. Majority of them distributed in water network regions, and lived with poor economic conditions. The average age of the patients was (66.3 +/- 9.3), with 89.3% ranged from 50 to 80 years old. Clinically 71.3% of the cases were with splenomegaly, 27.6% with ascites, 0.9% cases of multiple granuloma in the colon and 0.2% cases with dwarfism. 1 023 patients (96.5%) had received medical treatments. 69.4% of the cases had serious complications with advanced schistosomiasis and 52.5% had concurrently disorders in other systems. 71.1% of the patients had subjective symptoms and 65.2% had hepatosplenomegaly with hepatic fibrosis and dysfunction. The serum positive rate of anti-S. japonicum antibody was 15.7%. Stool hatching test and microscopy revealed no eggs in fecal samples, but 24 out of 38 cases were found metamorphic eggs by rectal biopsy. Three years' medical treatment improved the clinical conditions in 74.3% of the cases. CONCLUSION: Situation of the advanced schistosomiasis patients is quite critical. Treatment and care are urgently needed especially for those aged 50 to 80 years old. [

[Establishment and application of rapid dipstick of latex immunochromatographic assay for the diagnosis of schistosomiasis].

OBJECTIVE: To establish a simple and fast diagnostic assay for schistosomiasis. METHODS: Based on the immunochromatographic technique and the principle of indirect assay of ELISA, using soluble egg antigen (SEA) of Schistosoma japonicum and mouse anti-human monoclonal antibody labelled with red latex as color developing agents, a latex immunochromatographic assay (DLIA) was developed. Serum samples from 69 schistosomiasis patients were detected by DLIA. Tested were also 264 sera from healthy people, 15 sera from clonorchiasis patients, 8 sera from patients with angiostrongyliasis cantonensis, 11 sera from patients with intestinal nematode infection and 19 sera from paragonimiasis patients. ELISA was used as a parallel control. RESULTS: The sensitivity for detecting schistosomiasis antibodies with DLIA and ELISA was 94.2% (65/69) and 95.7% (66/69), respectively (chi2=0.15, P>0.05). The specificity in examining healthy persons was 97.4% (257/264) and 94.7% (250/264), respectively (chi2=2.43, P>0.05). No cross reaction was found with the sera of clonorchiasis, intestinal nematode infection and angiostrongyliasis. The cross reaction rate with paragonimiasis of the two assays was 42.1% (8/19) and 47.4% (9/19), respectively (chi2=0.11, P>0.05). CONCLUSION: DLIA is a simple, fast, sensitive and specific assay for the diagnosis of schistosomiasis.

A randomized, double-blind, multicenter clinical trial on the efficacy of ivermectin against intestinal nematode infections in China.

To assess the efficacy of ivermectin against intestinal nematode infections, a randomized, double-blind, multicenter clinical trial was carried out in a total of 816 human individuals infected with different nematodes from three counties in China. The subjects were randomly assigned into experimental and control groups and orally given a single dose of 0.1, 0.2, 0.2 and 0.2mg/kg ivermectin against Ascaris lumbricoides, hookworm, Trichuris trichiura and Enterobius vermicularis, respectively. Parallel control groups to each of the ivermectin groups were given a single oral dose of 6.7 mg/kg albendazole. The cure rates with ivermectin and albendazole were 100% (102/102) and 99.0% (101/102) for Ascaris, and 66.7% (68/102) and 67.7% (69/102) for Trichuris, respectively, with no significant difference (P>0.05) between the two treatments. The parasitological cure rates of albendazole were 69.6% (71/102) for hookworm and 94.1% (96/102) for Enterobius, which were significantly higher than ivermectin (33.3% and 52.9%, respectively, P<0.0001). The expulsion of worm in the feces reached its peak 1-2 days after ivermectin treatment. The study showed that ivermectin, with few side effects, could be used as an additional treatment tool for intestinal nematodes, especially for the treatment of Ascaris and Trichuris infections in China.

[Observation on the change of anti-S. japonicum antibody level in population migrated from outside embankment to new town].

OBJECTIVE: To detect the change of the anti-S. japonicum antibody level after people migrated from outside embankment to newly established town. METHODS: Three pilot spots were established for the investigation: one spot that both inhabitancy and cultivation disuse (A), one spot that only inhabitancy disuse but farming continued (B) and the third one served as control (C). DIGFA and ELISA were used to detect the antibody level in the populations from 2002 to 2005. RESULTS: The positive rate of anti-S. japonicum antibody declined significantly from 6.63% to 3.52% by DIGFA and from 7.26% to 3.71% by ELISA at spot A (chi2=5.2625, P<0.05; chi2=6.3296, P<0.05, respectively). There was no significant difference on the positive rate of antibody in spots B and C. The average A450 value of ELISA in the three spots was statistically analyzed by One-Way ANOVA. It was only in spot B that the average A450 value declined from 0.182 in 2003 to 0.147 in 2005 (P<0.01). CONCLUSION: The anti-S. japonicum antibody level in human population has decreased at certain degree after they migrated from outside embankment to new town.

[Prophylactic effect of artesunate against experimental infection of Schistosoma mansoni].

OBJECTIVE: To study the prophylactic effect of artesunate against the infection of Schistosoma mansoni in mice and its optimal scheme for preventing schistosomiasis mansoni. METHODS: BALB/c mice were infected by tail dipping method with S. mansoni cercariae. Mice were administered orally with artesunate at different developmental stage of the parasite, with different regimens. The reduction rates of total and female worms, the number of eggs in the liver and intestine, and the fecundity were calculated and treated statistically. RESULTS: The optimal dosage of artesunate to prevent murine schistosomiasis was 300 mg/kg. The parasite was found to be especially susceptible to artesunate in its schistosomula stage of 14 and 21 d after infection, resulting in worm reduction rate of 84% and 93% respectively compared with control. High protection was reached with worm reduction rate of 99% by the regimens of 300 mg/kg once a week for 4 consecutive weeks beginning 14 d after infection. The fecundity was significantly suppressed, suggesting that the drug inhibited sexual maturation of female worms. The effective protection could also be gained with prolonged interval time of two weeks with worm reduction rate of 97% and 96% beginning 14 or 21 d after infection. CONCLUSION: Artesunate kills schistosomula and reduces the fecundity of females effectively, the infected mice do not develop schistosomiasis mansoni when treated with artesunate. It's proposed that an optimal scheme for field use be the first administration 14 or 21 days after infection with 1 or 2 weeks interval.

[Clinical observation on the efficacy of ivermectin in the treatment of intestinal nematode infections].

OBJECTIVE: To observe the efficacy and side effects of ivermectin in the treatment of intestinal nematode infections. METHODS: A single dose of ivermectin 0.1, 0.2, 0.2 and 0.2 mg/kg was orally administered to cases with infection of Ascaris, hookworm, Trichuris and Enterobius respectively. A single dose of albendazole 400 mg was used as control. RESULTS: The egg negative conversion rates of ivermectin and albendazole were both 100% (34/34) for Ascaris infection, 17.6% (6/34) and 76.5% (26/34) respectively for hookworm infection, 67.6% (23/34) and 47.1% (16/34) respectively for Trichuris infection, 58.8% (20/34) and 100% (34/34) respectively for Enterobius infection. The worm discharge reached a peak in 1-2 days after treatment. The side effect of ivermectin was mild and transient showing no adverse effect on blood picture, liver function, renal function or ECG. CONCLUSION: Ivermectin shows similar effect on Ascaris with albendazole, better effect on Trichuris and poorer effect on hookworm and Enterobius than albendazole.