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Blastocystis is a common intestinal protist in humans and animals. Currently, Blastocystis infection in bar-headed geese is unknown. To understand the prevalence and distribution of Blastocystis subtypes in bar-headed geese, fecal samples were collected from 358 bar-headed geese in three regions. The total infection rate of Blastocystis in bar-headed in China was 5.9% (21/358), with 5.0% (6/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 11.8% (14/119) in Maqu county, Gansu province, and 0.8% (1/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rates by region were statistically significant. Sequences analyses showed that one known subtype (ST7, n = 16) and one potentially novel ST (n = 5) in bar-headed geese were detected in this study. This is the first report on the prevalence and subtype distribution of Blastocystis in bar-headed geese, which will improve our understanding of the epidemiology and public health implications of Blastocystis infection in wild migratory birds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Cryptosporidium spp. is an important foodborne and waterborne pathogen in humans and animals, causing diarrhoea in humans and respiratory and gastrointestinal diseases in birds. However, reports of Cryptosporidium infection in bar-headed goose are limited. To determine the infection rate and species/genotypes of Cryptosporidium in bar-headed goose in China, a total of 358 fecal samples were collected from 3 regions. Nested PCR was used to amplify Cryptosporidium SSU rRNA regions from the fecal extracted-DNA samples. The total infection rate of Cryptosporidium in bar-headed in China was 3.9 % (14/358), with 4.2 % (5/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 7.6 % (9/119) in Maqu county, Gansu province, and 0.0 % (0/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rate by region were statistically significant. All positive samples were identified as Cryptosporidium goose genotype I (n = 14). This is the first systematic investigation of the epidemiological status and dominant species/genotypes of Cryptosporidium in bar-headed goose in China, thereby enhancing our understanding of the epidemiology of Cryptosporidium infection in wild migratory birds., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Pathogens can impact host RNA modification machinery to establish a favorable cellular environment for their replication. In the present study, we investigated the effect of Toxoplasma gondii infection on host RNA modification profiles and explored how these modifications may influence the host-parasite interaction., Methodology/principal Findings: We analyzed the modification levels of ∼ 80 nt tRNA and 17-50 nt sncRNAs in mouse liver, spleen, and serum using liquid chromatography and tandem mass spectrometry analysis. The results revealed alterations in RNA modification profiles, particularly during acute infection. The liver exhibited more differentially abundant RNA modifications than the spleen. RNA modification levels in serum were mostly downregulated during acute infection compared to control mice. Correlations were detected between different RNA modifications in the liver and spleen during infection and between several RNA modifications and many cytokines. Alterations in RNA modifications affected tRNA stability and protein translation., Conclusions/significance: These findings provide new insight into the role of RNA modifications in mediating the murine host response to T. gondii infection., Competing Interests: The authors declare that there are no competing interests., (Copyright: © 2024 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Understanding the characteristics of bacteriophages is crucial for the optimization of phage therapy. In this study, the biological and genomic characteristics of coliphage LHE83 were determined and its synergistic effects with different types of antibiotics against E. coli E82 were investigated. Phage LHE83 displayed a contractile tail morphology and had a titer of 3.02 × 10 9 pfu/mL at an optimal MOI of 0.01. Meanwhile, phage LHE83 exhibited good physical and chemical factors tolerance. The 1-step growth analysis revealed a latent period of approx. 10 min with a burst size of 87 pfu/infected cell. Phage LHE83 belongs to the genus Dhakavirus. Its genome consists of 170,464 bp with a 40% GC content, and a total of 268 Open Reading Frames (ORF) were predicted with no detected virulent or resistant genes. ORF 213 was predicted to encode the receptor binding protein (RBP) and confirmed by the antibody-blocking assay. Furthermore, a phage-resistant strain E. coli E82R was generated by co-culturing phage LHE83 with E. coli E82. Genomic analysis revealed that OmpA served as the receptor for phage LHE83, which was further confirmed by phage adsorption assay using E. coli BL21ΔOmpA, E. coli BL21ΔOmpA: OmpA and E. coli BL21:OmpA strains. Additionally, a synergistic effect was observed between phage LHE83 and spectinomycin against the drug-resistant strain E. coli E82. These results provide a theoretical basis for understanding the interactions between phages, antibiotics, and host bacteria, which can assist in the clinical application of phages and antibiotics against drug-resistant bacteria., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Blastocystis is a protist that is distributed in the gut tract of humans and animals. However, the reports about Blastocystis infection in Tibetan antelope are scarce. We collected 173 Tibetan antelope feces samples from Xinjiang, Qinghai and Xizang, and amplified the SSU rRNA gene of 600 bp region of Blastocystis in our research. Fifty-one samples in total were positive for Blastocystis, with all subtypes being ST31. The lowest prevalence of Blastocystis was observed in Xizang (2/20, 9.1%), followed by Qinghai (18/92, 16.4%), Xinjiang (31/61, 33.7%). The highest prevalence of Blastocystis in Tibetan antelope was detected during the summer was (19/30, 38.8%). This is the first research work regarding the Blastocystis subtypes ST31 in Tibetan antelope. Our research provides information for future researches on the distribution of this Blastocystis subtype and the control of Blastocystis infection., Competing Interests: Declaration of competing interest The authors confirm that they have no known financial or interpersonal conflicts that would have appeared to have an impact on the research presented in this study., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Avian malaria is a vector-borne parasitic disease caused by Plasmodium infection transmitted to birds by mosquitoes. The aim of this systematic review was to analyze the global prevalence of malaria and risk factors associated with infection in wild birds. A systematic search of the databases CNKI, WanFang, VIP, PubMed, and ScienceDirect was performed from database inception to 24 February 2023. The search identified 3181 retrieved articles, of which 52 articles met predetermined inclusion criteria. Meta-analysis was performed using the random-effects model. The estimated pooled global prevalence of Plasmodium infection in wild birds was 16%. Sub-group analysis showed that the highest prevalence was associated with adult birds, migrant birds, North America, tropical rainforest climate, birds captured by mist nets, detection of infection by microscopy, medium quality studies, and studies published after 2016. Our study highlights the need for more understanding of Plasmodium prevalence in wild birds and identifying risk factors associated with infection to inform future infection control measures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Enterocytozoon bieneusi, a zoonotic pathogen prevalent in both humans and animals, is the most frequently diagnosed microsporidian species in humans and presents significant public health risks. However, data on the prevalence and genotypes of E. bieneusi in farmed minks (Neovison vison) and raccoon dogs (Nyctereutes procyonoides) in China are limited. Therefore, 275 minks (89 from Hebei Province, 57 from Heilongjiang Province, 109 from Liaoning Province, 20 from Shandong Province) and 235 raccoon dogs (114 from Hebei Province, 27 from Heilongjiang Province, 61 from Liaoning Province, 33 from Jilin Province) were examined for the prevalence and genotypes of E. bieneusi through sequence analysis of the internal transcribed spacer (ITS) region of the rRNA gene. The overall prevalence of E. bieneusi was 18.6% (95/510), with 10.5% (29/275) in farmed minks and 28.1% (66/235) in raccoon dogs. Ten genotypes (CHN-F1, genotype D, Type IV, EbpC, NCF2, NCF5, NCF6, Peru8, Henan V, and MJ5) were identified in minks and raccoon dogs. This study is the first to detect the CHN-F1, NCF2, NCF6, Peru8, and Henan V genotypes in minks and the NCF5, NCF6, and MJ5 genotypes in raccoon dogs. Additionally, the D, Type IV, and Peru8 genotypes, previously identified in humans, were also found in minks and raccoon dogs, suggesting that these animals could be potential sources of human microsporidiosis. These findings expand the understanding of E. bieneusi's host distribution in China and contribute to the prevention of zoonotic E. bieneusi infections among farmed animals., (© N.-Y. Xue et al., published by EDP Sciences, 2024.)

Background: The gut microbiota plays an essential role in maintaining gut homeostasis and improving performance, with the composition of microbial communities visibly differing across different laying stages in hens and significantly correlating with egg production. To gain further insights into the association between microbial community characteristics and laying periods in Hy-Line variety brown and Isa brown laying hens, we conducted a 16S rRNA amplicon sequencing survey., Results: Our result revealed the diversity of bacteria in the early laying period was commonly higher than peak, and in Hy-Line variety brown laying hens were generally higher than Isa brown. Principal coordinate analysis (PCoA) and permutational multivariate analysis of variance (PERMANOVA) revealed that the structure and composition of the gut microbiota of laying hens exhibited significant differences among different groups. Phylum Firmicutes, Bacteroidota, Proteobacteria, and Fusobacteriota were found that dominant in the host's feces. Therein, the abundance of Fusobacteriota was higher in the peak period than in the early period, while the abundance of Cyanobacteria in the early period was higher in two breeds of hens. Furthermore, random forest based on machine learning showed that there were several distinctly abundant genera, which can be used as potential biomarkers to differentiate the different groups of laying periods and breeds. In addition, the prediction of biological function indicated the existing discrepancy in microbial function among the microbiota of four groups., Conclusions: Our findings offer new insights into the bacterial diversity and intestinal flora composition of different strains of laying hens during various laying periods, contributing significantly to the improvement of production performance and the prevention of chicken diseases., (© 2023. The Author(s).)

Bovine coronavirus (BCoV) is one of the important pathogens that cause calf diarrhea (CD), winter dysentery (WD), and the bovine respiratory disease complex (BRDC), and spreads worldwide. An infection of BCoV in cattle can lead to death of young animals, stunted growth, reduced milk production, and milk quality, thus bringing serious economic losses to the bovine industry. Therefore, it is necessary to prevent and control the spread of BCoV. Here, a systematic review and meta-analysis was conducted to assess the prevalence of BCoV in cattle in China before 2022. A total of 57 articles regarding the prevalence of BCoV in cattle in China were collected from five databases (PubMed, ScienceDirect, CNKI, VIP, and Wan Fang). Based on the inclusion criteria, a total of 15,838 samples were included, and 6,136 were positive cases. The overall prevalence of BCoV was 30.8%, with the highest prevalence rate (60.5%) identified in South China and the lowest prevalence (15.6%) identified in Central China. We also analyzed other subgroup information, included sampling years, sample sources, detection methods, breeding methods, age, type of cattle, presence of diarrhea, and geographic and climatic factors. The results indicated that BCoV was widely prevalent in China. Among all subgroups, the sample sources, detection methods, breeding methods, and presence or absence of diarrheal might be potential risk factors responsible for BCoV prevalence. It is recommended to strengthen the detection of BCoV in cattle, in order to effectively control the spread of BCoV., Competing Interests: Declaration of competing interest The authors have no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The fungal community, also known as mycobiota, plays pivotal roles in host nutrition and metabolism and has potential to cause disease. However, knowledge of the gut fungal structure in Caprinae is quite limited. In this study, the composition and diversity of the gut mycobiota of Caprinae animals from different geographical locations (Anhui, Jilin, Guangxi, Shandong, Shanxi, and Tibet) were comprehensively characterized by analyzing the internal transcribed spacer 2 (ITS-2) sequences of the fungal community. The results showed that Ascomycota and Basidiomycota were the dominant phyla, which, respectively, accounted for 90.86 to 95.27% and 2.58 to 7.62% of sequences in samples from each region. Nonetheless, the structure of the gut mycobiota was largely different in Caprinae animals in the different provinces. Therein, Sporormiaceae and Thelebolaceae were the dominant fungal families in the samples from Tibet, whereas their abundance was generally low in other regions. The intestinal diversity of individuals from Guangxi was higher than that in other regions. In addition, there were 114 differential genera among all regions. Finally, the co-occurrence network revealed 285 significant correlations in cross-family pairs in the guts of Caprinae animals, which contained 149 positive and 136 negative relationships, with 96 bacterial and 86 fungal participants at the family level. This study has improved the understanding of the mycobiota of ruminants and provided support for the improvement in animal health and productivity. IMPORTANCE In this study, we elucidated and analyzed the structure of the gut mycobiota of Caprinae animals from different regions. This study revealed differences in the structure of the gut mycobiota among Caprinae animals from different geographical environments. Based on previous findings, correlations between fungal and bacterial communities were analyzed. This study adds to previous research that has expanded the present understanding of the gut microbiome of Caprinae animals.

Most microbiome studies regarding the ruminant digestive tract have focused on the rumen microbiota, whereas only a few studies were performed on investigating the gut microbiota of ruminants, which limits our understanding of this important component. Herein, the gut microbiota of 30 Caprinae animals (sheep and goats) from six provinces in China was characterized using ultradeep (>100 Gbp per sample) metagenome shotgun sequencing. An inventory of Caprinae gut microbial species containing 5,046 metagenomic assembly genomes (MAGs) was constructed. Particularly, 2,530 of the genomes belonged to uncultured candidate species. These genomes largely expanded the genomic repository of the current microbes in the Caprinae gut. Several enzymes and biosynthetic gene clusters encoded by these Caprinae gut species were identified. In summary, our study extends the gut microbiota characteristics of Caprinae and provides a basis for future studies on animal production and animal health. IMPORTANCE We constructed a microbiota catalog containing 5,046 MAGs from Caprinae gut from six regions of China. Most of the MAGs do not overlap known databases and appear to be potentially new species. We also characterized the functional spectrum of these MAGs and analyzed the differences between different regions. Our study enriches the understanding of taxonomic, functional, and metabolic diversity of Caprinae gut microbiota. We are confident that the manuscript will be of utmost interest to a wide range of readers and be widely applied in future research.

Echinococcosis is a foodborne parasitic zoonosis caused by the larvae of Echinococcus . This disease can affect goats and other mammals. In this study, a systematic review and meta-analysis for echinococcosis in global goats were performed based on the following five databases (China National Knowledge Infrastructure [CNKI], VIP Chinese Journal Database, Wanfang Data, PubMed, and ScienceDirect). In total, 108,197 samples were collected. The global prevalence of echinococcosis in goats was identified to be 10.85% (3217/108,197). The prevalence of echinococcosis in goats was 6.16% (1369/22,208) and 13.27% (874/5932) in South America and Africa, respectively. The prevalence of echinococcosis in goats before 2010 (9.76%; 112/713) was significantly higher than that from 2010 to 2014 (1.44%; 45/32,145) or after 2014 (2.95%; 154/3889). The prevalence of echinococcosis in goats aged <12 months (4.48%; 70/2911) was higher than that in goats aged ≥12 months (2.88%; 36/819). We also investigated the effects of geographical factors and climates on the prevalence of echinococcosis in goats. The results showed that the prevalence of echinococcosis was higher in the areas with high altitude and cold climate. This meta-analysis indicated that echinococcosis was ubiquitous in goats. Thus, we should improve the feeding conditions for goats, and strengthen the control measures of echinococcosis epidemic in goats, with the aims of reducing the economic losses of animal husbandry and providing protection for humans in the aspects of food security and health.

Background: Portal hypertension monitoring is important throughout the natural course of cirrhosis. Hepatic venous pressure gradient (HVPG), regarded as the golden standard, is limited by invasiveness and technical difficulties. Portal hypertension is increasingly being assessed non-invasively, and hematological indices, imaging data, and statistical or computational models are studied to surrogate HVPG. This paper discusses the existing non-invasive methods based on measurement principles and reviews the methodological developments in the last 20 years., Methods: First, we used VOSviewer to learn the architecture of this field. The publications about the non-invasive assessment of portal hypertension were retrieved from the Web of Science Core Collection (WoSCC). VOSviewer 1.6.17.0 was used to analyze and visualize these publications, including the annual trend, the study hotspots, the significant articles, authors, journals, and organizations in this field. Next, according to the cluster analysis result of the keywords, we further retrieved and classified the related studies to discuss., Results: A total of 1,088 articles or review articles about our topic were retrieved from WoSCC. From 2000 to 2022, the number of publications is generally growing. "World Journal of Gastroenterology" published the most articles ( n = 43), while "Journal of Hepatology" had the highest citations. "Liver fibrosis" published in 2005 was the most influential manuscript. Among the 20,558 cited references of 1,088 retrieved manuscripts, the most cited was a study on liver stiffness measurement from 2007. The highest-yielding country was the United States, followed by China and Italy. "Berzigotti, Annalisa" was the most prolific author and had the most cooperation partners. Four study directions emerged from the keyword clustering: (1) the evaluation based on fibrosis; (2) the evaluation based on hemodynamic factors; (3) the evaluation through elastography; and (4) the evaluation of variceal bleeding., Conclusion: The non-invasive assessment of portal hypertension is mainly based on two principles: fibrosis and hemodynamics. Liver fibrosis is the major initiator of cirrhotic PH, while hemodynamic factors reflect secondary alteration of splanchnic blood flow. Blood tests, US (including DUS and CEUS), CT, and magnetic resonance imaging (MRI) support the non-invasive assessment of PH by providing both hemodynamic and fibrotic information. Elastography, mainly USE, is the most important method of PH monitoring., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sun, Ni, Xue, Wang, Aljbri, Wang, Ren, Li and Niu.)

The diversity of bacteria and fungi in the gut microbiota of commercial broilers that raised in cages from hatch to the end of the production cycle were examined by an analysis of 3,592 and 3,899 amplicon sequence variants (ASVs), respectively. More than 90% sequences in bacterial communities were related to Firmicutes and Proteobacteria. More than 90% sequences in fungal communities were related to Ascomycota, Basidiomycota, and Glomeromycota. A statistical analysis of the microbiota composition succession showed that age was one of the main factors affecting the intestinal microbial communities of broilers. The increasingly complex community succession of transient microbiota occurred along with an increase of age. This dynamic change was observed to be similar between bacteria and fungi. The gut microbiota had a special structure in the first 3 d after birth of broiler. The microbiota structure was quite stable in the period of rapid skeletal growth (d 14-21), and then changed significantly in the period of rapid gaining weight (d 35-42), thus indicating the composition of gut microbiota in broilers had unique structures at different developmental stages. We observed that several bacteria and fungi occupied key functions in the gut microbiota of broilers, suggesting that the gut homeostasis of broilers might be affected by losses of bacteria and fungi via altering interactions between microbiota. This study aimed to provide a data basis for manipulating the microbiota at different developmental stages, in order to improve production and the intestinal health of broilers., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Toxoplasma gondii ( T. gondii ) infection can cause intestinal inflammation in rodents and significantly alters the structure of gut microbiota. However, the effects of different T. gondii genotypes on the gut microbiota of rats remain unclear. In this study, acute and chronic T. gondii infection in Fischer 344 rats was induced artificially by intraperitoneal injection of tachyzoites PYS (Chinese 1 ToxoDB#9) and PRU (Type II). Fecal 16S rRNA gene amplicon sequencing was employed to analyze the gut microbiota structure at different stages of infection, and to compare the effects of infection by two T. gondii genotypes. Our results suggested that the infection led to structural changes of gut microbiota in rats. At the acute infection stage, the microbiota diversity increased, while both diversity and abundance of beneficial bacteria decreased at the chronic infection stage. The differences of microbiota structure were caused by strains of different genotypes. However, the diversity changes were consistent. This study demonstrates that the gut microbiota plays an important role in T. gondii infection in rats. The data will improve our understanding of the association between T. gondii infection and gut microbiota in rodents., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lv, Ma, Wei, Qin, Qiu, Ni, Yang and Cao.)

Background: The Delta COVID-19 variant caused a resurgence in cases and deaths during the summer of 2021, particularly among the unvaccinated, highlighting the need to increase vaccine coverage. We describe a survey conducted in September 2021, in the midst of the Delta variant surge, after the FDA fully approved Pfizer-BioNTech’s vaccine for ages 16+ and issued an emergency use authorization for ages 12−15. Methods and Findings: US adults were surveyed to measure COVID-19 vaccination status, intentions, attitudes, values, and trust in public health authorities. More than three-quarters (77%) reported receiving at least one dose of COVID-19 vaccination. Of the unvaccinated, 6% intended to vaccinate, 40% were unlikely to ever vaccinate, and 55% remained uncertain. Most of the unvaccinated were <45 years old (62%), without a bachelor’s degree (83%), earning less than $85,000 annually (74%), and Republican/Independent (66%). Concerns among the unvaccinated-yet-still-uncertain included the vaccines’ safety (86%), speed of development (86%), and suspicion of government (79%) and pharmaceutical companies (69%). Most (86%) of the unvaccinated reported they would not vaccinate if mandated by their employer. About one third (34%) of the unvaccinated reported facing at least one barrier to vaccination. Conclusion: More than half of unvaccinated adults remained uncertain about COVID-19 vaccination, indicating an opportunity to support their decision making. Public health must increase easy and equitable access to vaccination and renew efforts to provide unvaccinated populations access to information from trusted sources.

Blastocystis is a common unicellular protist that lives in the intestines of humans and animals. Blastocystis infection and subtypes in cattle have been reported in several regions. However, the information of Blastocystis infection in cattle in China is still largely scant. To assess the prevalence and subtype distribution of Blastocystis in beef cattle in China, 803 fecal samples were collected from beef cattle farms in four cities of Northeast China, and were subjected to an analysis based on small subunit rRNA gene fragment. The overall prevalence of Blastocystis in beef cattle was 2.11% (17/803), with 2.15% in preweaning calves, 1.9% in postweaning calves, and 3.85% in breeding cattle, but absence in adult cattle ( p  > 0.05). Moreover, five Blastocystis subtypes were identified (ST10, ST21, ST23, ST25, and ST26), among which ST10 and ST26 subtypes were dominant subtypes in beef cattle. Mixed infections were detected in three specimens (ST10/ST25, ST10/ST23/ST25, and ST10/ST26). This is the first report showing Blastocystis infection in beef cattle in Northeast China. In addition, a variety of Blastocystis subtypes are reported in cattle in China for the first time. These results will benefit for better understanding the epidemiology and public health implications of Blastocystis.

Enterocytozoon ( E. ) bieneusi and Cryptosporidium spp. are the most important zoonotic enteric pathogens associated with diarrheal diseases in animals and humans. However, it is still not known whether E. bieneusi and Cryptosporidium spp. are carried by wild rodents in Shanxi, Guangxi, Zhejiang, Shandong, and Inner Mongolia, China. In the present study, a total of 536 feces samples were collected from Rattus (R.) norvegicus , Mus musculus , Spermophilus ( S. ) dauricus , and Lasiopodomys brandti in six provinces of China, and were detected by PCR amplification of the SSU rRNA gene of Cryptosporidium spp. and ITS gene of E. bieneusi from June 2017 to November 2020. Among 536 wild rodents, 62 (11.6%) and 18 (3.4%) samples were detected as E. bieneusi - and Cryptosporidium spp.-positive, respectively. Differential prevalence rates of E. bieneusi and Cryptosporidium spp. were found in different regions. E. bieneusi was more prevalent in R. norvegicus , whereas Cryptosporidium spp. was more frequently identified in S. dauricus . Sequence analysis indicated that three known Cryptosporidium species/genotypes ( Cryptosporidium viatorum , Cryptosporidium felis , and Cryptosporidium sp. rat genotype II/III) and two uncertain Cryptosporidium species ( Cryptosporidium sp. novel1 and Cryptosporidium sp. novel2) were present in the investigated wild rodents. Meanwhile, 5 known E. bieneusi genotypes (XJP-II, EbpC, EbpA, D, and NCF7) and 11 novel E. bieneusi genotypes (ZJR1 to ZJR7, GXM1, HLJC1, HLJC2, and SDR1) were also observed. This is the first report for existence of E. bieneusi and Cryptosporidium spp. in wild rodents in Shanxi, Guangxi, Zhejiang, and Shandong, China. The present study also demonstrated the existence of E. bieneusi and Cryptosporidium spp. in S. dauricus worldwide for the first time. This study not only provided the basic data for the distribution of E. bieneusi and Cryptosporidium genotypes/species, but also expanded the host range of the two parasites. Moreover, the zoonotic E. bieneusi and Cryptosporidium species/genotypes were identified in the present study, suggesting wild rodents are a potential source of human infections., Competing Interests: Author Y-CW is employed by Veterinary Department, Muyuan Foods Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ni, Sun, Qin, Wang, Zhao, Sun, Zhang, Yang, Feng, Guan, Qiu, Wang, Xue and Sun.)

Cryptosporidium spp., the causative agent of cryptosporidiosis, can infect a variety of hosts. So far, there has been limited information regarding Cryptosporidium spp. infection in yaks ( Bos grunniens ). Here, we performed the first systematic review and meta-analysis for Cryptosporidium spp. infection in yaks in China. To perform the meta-analysis, five databases (Chinese National Knowledge Infrastructure (CNKI), VIP Chinese journal database, WanFang Data, PubMed, and ScienceDirect) were employed to search for studies related to the prevalence of Cryptosporidium spp. in yaks in China. The total number of samples was 8,212, and the pooled Cryptosporidium spp. prevalence in yaks was estimated to be 10.52% (1192/8012). The prevalence of Cryptosporidium spp. in yaks was 13.54% (1029/5277) and 4.49% (148/2132) in northwestern and southwestern China, respectively. In the sampling year subgroups, the prevalence before 2012 (19.79%; 650/2662) was significantly higher than that after 2012 (6.07%; 437/4476). The prevalence of Cryptosporidium spp. in cold seasons (20.55%; 188/794) was higher than that in warm seasons (4.83%; 41/1228). In the age subgroup, the yaks with age < 12 months had a higher prevalence (19.47%; 231/1761) than that in yaks with age ≥12 months (16.63%; 365/2268). Among 12 Cryptosporidium spp. species/genotypes, the C. bovis had the highest prevalence. Moreover, the effects of geography (latitude, longitude, precipitation, temperature, and altitude) and climate on Cryptosporidium spp. infection in yaks were evaluated. Through analyzing the risk factors correlated with the prevalence of Cryptosporidium spp., we recommend that effective management measures should be formulated according to the differences of different geographical factors, in order to prevent cryptosporidiosis and reduce economic losses in yaks in China., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Geng, Ni, Li, Jiang, Wang, Wei, Zhang and Sun.)

Blastocystis is a protozoan that parasitizes the intestines. A number of hosts of Blastocystis have been found, including human and animals. However, there has been no research on the prevalence of Blastocystis in Tibetan antelope. Here, a molecular test was performed using 627 Tibetan antelope fecal samples collected on Tibet in China from 2019 to 2020. The result showed that 30 (4.8%) samples were Blastocystis positive. The highest prevalence of Blastocystis was in Shuanghu County (25/209, 12.0%), followed by Shenza County (2/103, 1.9%), Nyima County (3/182, 1.6%), and Baigoin County (0/133, 0.0%). In addition, logistic regression analysis showed that the gender, sampling year, and area of Tibetan antelope were risk factors for Blastocystis prevalence. Three subtypes (ST10, ST13, and ST14) of Blastocystis were found in Tibetan antelope through a subtype sequence analysis, and ST13 was identified to be the dominant subtype. This is the first investigation for the infection of Blastocystis in Tibetan antelope. Collectively, the data in this study have expanded the host range of Blastocystis and provided basic information for the distribution of Blastocystis subtypes, which could support the prevention of Blastocystis infection in wild animals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Geng, Sun, Jiang, Sun, Li, Qin, Wang, Ma, Zhu, Xue and Ni.)

Toxoplasmosis is a zoonotic disease caused by Toxoplasma gondii, which is widespread in warm-blooded animals and humans. Currently, many studies regarding T. gondii infection in sheep have been published worldwide. However, there is no meta-analysis of sheep infection in China. In this study, five databases were used to retrieve articles related to T. gondii in China. A total of 59 studies from 1987 to June 30, 2020, have been included. We estimated that the combined prevalence of T. gondii in the selected period was 8.5% (3197/28,099). In the analysis of publication year, the lowest positive rate after 2010 was 7.4% (1883/17,313). Geographically, the highest prevalence of T. gondii in sheep was recorded in Southwest China 19.2% (490/2080). Analysis according to age showed that the prevalence of infection in sheep older than 12 months was significantly (P < 0.05) higher than that in younger animals. The prevalence of T. gondii in sheep farmed by farmers was 7.2% (566/6336), which was higher than 5.3% (660/3121) of large-scale farming, suggesting a role for the feeding and husbandry. We also analyzed the impact of different geographic and climatic factors on the prevalence of T. gondii infection in sheep. The results showed that the prevalence was higher in low altitude, warm, humid, and high rainfall areas. We suggest that appropriate control programs should be formulated according to the differences in reproduction patterns and geographical conditions in different regions to reduce the prevalence of T. gondii among sheep in China., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Cryptosporidium is an important zoonotic parasite that can infect a variety of hosts, including pigs and humans, through water and food. Many studies on Cryptosporidium infection in pigs have been reported worldwide. However, the meta-analysis of Cryptosporidium infection in pigs in China has not been published. This study retrieved articles related to Cryptosporidium in pigs in China by using four databases: Chinese National Knowledge Infrastructure (CNKI), PubMed, VIP Chinese journal database and Wanfang Data. We retrieved 40 studies related to Cryptosporidium infection in China, and those articles were harvested from the inception to 1 January 2020. We estimated that the overall prevalence of pigs with Cryptosporidium in the selected period was 12.2% (4,349/30,404). In the sampling year subgroup, the prevalence rate after 2010 was the lowest at 8.7% (2,087/18,100). In Northern China, the Cryptosporidium prevalence was 47.9% (34/71). By contrast, the prevalence of Cryptosporidium in Southwestern China was only 6.9% (778/6,445). The infection rate of Cryptosporidium in diarrhoea pigs of 15.6% (74/384) was higher than that in non-diarrhoea pigs at 10.8% (378/2,840). Among the four age groups, the prevalence of weaning pigs of 16.2% (530/3,243) was the highest, and the difference was significant (p < .05). The prevalence of Cryptosporidium in extensive farming was 25.7% (660/3,121), which was significantly higher than in intensive farming 8.7% (566/6,336), and the prevalence of infection was related to the farming modes (p < .05). We also analysed the impact of different geographic factor subgroups (longitude, latitude, precipitation, temperature, humidity, climate and altitude) on the prevalence of pigs. The results showed that cryptosporidiosis was widespread in pigs in China. We suggest that appropriate control schemes should be developed according to the differences in breeding patterns and geographic conditions in different regions, and effective management measures should be developed to reduce the spread between pigs., (© 2020 Wiley-VCH GmbH.)

Protists of the Blastocystis genus are distributed worldwide and can infect a range of hosts. However, data concerning Blastocystis infection are limited for sika deer and are not available for black bears. Therefore, in the present study, a total of 312 black bears (Ursus thibetanus) from Heilongjiang Province and 760 sika deer (Cervus nippon) from four different northern Chinese provinces were investigated. Blastocystis infection in these animals was detected via PCR amplification of the small subunit rRNA gene in fecal samples. The prevalence of Blastocystis infection in black bears and sika deer was 14.4% (45/312 positive samples) and 0.8% (6/760 positive samples), respectively. Young black bears (18.3%) had a significantly higher Blastocystis prevalence than adult bears (9.1%). The prevalence of Blastocystis was significantly higher in black bears raised outdoors (24.6%) than in bears raised indoors (12.2%). Blastocystis-positive sika deer were only found in Jilin Province (1.3%, 6/480). Female sika deer (0%, 0/61) had a significantly lower Blastocystis prevalence than males (0.9%, 6/699). Sanger sequencing was used to determine the small subunit rRNA gene sequences of the Blastocystis-positive PCR products. A neighbor-joining phylogenetic tree based on the small subunit rRNA gene sequences showed that only Blastocystis subtype (ST)1 was identified in black bears, whereas ST10 and ST14 were found in sika deer. This is the first report of Blastocystis ST1 infection in black bears. These findings also extend the distribution information of Blastocystis subtypes, which will provide a foundation for further study of Blastocystis in different hosts in China.

Background: DNA vaccine is one of the research hotspots in veterinary vaccine development. Several advantages, such as cost-effectiveness, ease of design and production, good biocompatibility of plasmid DNA, attractive biosafety, and DNA stability, are found in DNA vaccines., Methods: In this study, the plasmids expressing bovine herpesvirus 1 (BoHV-1) gB, gC, and gD proteins were mixed at the same mass ratio and adsorbed polyethyleneimine (PEI) magnetic beads with a diameter of 50 nm. Further, the plasmid and PEI magnetic bead polymers were packaged into double carboxyl polyethylene glycol (PEG) 600 to use as a DNA vaccine. The prepared DNA vaccine was employed to vaccinate mice via the intranasal route. The immune responses were evaluated in mice after vaccination., Results: The expression of viral proteins could be largely detected in the lung and rarely in the spleen of mice subjected to a vaccination. The examination of biochemical indicators, anal temperature, and histology indicated that the DNA vaccine was safe in vivo. However, short-time toxicity was observed. The total antibody detected with ELISA in vaccinated mice showed a higher level than PBS, DNA, PEI + DNA, and PBS groups. The antibody level was significantly elevated at the 15th week and started to decrease since the 17th week. The neutralizing antibody titer was significantly higher in DNA vaccine than naked DNA vaccinated animals. The total IgA level was much greater in the DNA vaccine group compared to other component vaccinated groups. The examination of cellular cytokines and the percentage of CD4/CD8 indicated that the prepared DNA vaccine induced a strong cellular immunity., Conclusion: The mixed application of plasmids expressing BoHV-1 gB/gC/gD proteins by nano-carrier through intranasal route could effectively activate long-term humoral, cellular, and mucosal immune responses at high levels in mice. These data indicate PEI magnetic beads combining with PEG600 are an efficient vector for plasmid DNA to deliver intranasally as a DNA vaccine candidate.

Hepatitis E virus (HEV) is a zoonotic pathogen that has spread worldwide. The HEV reservoir associated with livestock hepatitis E poses a huge threat to public health. Awareness of the prevalence and spatial distribution of livestock hepatitis E is valuable to prevent and control diseases caused by HEV, especially human hepatitis E infection. Currently, swine, including pigs (Sus scrofa), are recognized as the major reservoir of HEV. Therefore, we conducted a systematic review and meta-analysis to evaluate the pooled prevalence of HEV among swine in China. A total of 71 published papers on HEV infection in swine in China (including data from 49,523 animals) from January 1, 2010 to December 31, 2019 met the standard after searching five databases including the Technology Periodical Database, the Wan Fang Database, the China National Knowledge Infrastructure, PubMed, and ScienceDirect. A random effects model was used to calculate the pooled prevalence of HEV in swine. The results showed that the seroprevalence was 48.0% (95% confidence interval (CI) 39.6-56.9) and the prevalence of HEV RNA was 14.4% (95% CI 10.7-18.5). The estimated overall prevalence was 34.1% (95% CI 27.2-41.4). Central China (68.0%, 95% CI 42.2-89.1) had a significantly higher prevalence than other regions. In the publication year subgroup, the prevalence in 2016 or later (27.2%, 95% CI 19.3-36.0) was significantly lower than that in 2011 or earlier (49.0%, 95% CI 36.2-61.8). The prevalence of IgG (42.9%, 95% CI 31.7-54.6) was significantly higher than that of IgM (4.9%, 95% CI 1.6-9.7). Suckling piglets (15.6%, 95% CI 6.6-27.1) had a lower prevalence compared with that in other age groups. In all sample types, body fluids showed the highest prevalence (50.5%, 95% CI 41.7-59.3). Moreover, the pooled prevalence of HEV in boars was higher than that in sows (35.4% > 17.3%). The analysis suggested that HEV infection is common among swine in China. Further strengthening HEV testing in boars, controlling environmental pollution, and reducing the mixed feeding of different stages could contribute to reducing HEV infection in pigs in China and the risk of porcine HEV infection in humans., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can cause diarrhea and other gastrointestinal illnesses in humans and animals. However, the prevalence and genotype of the parasites in Longjiang Wagyu cattle in Heilongjiang Province, Northeast China have not been reported. In the present study, a total of 423 fecal samples of Longjiang Wagyu cattle collected from different farms in Heilongjiang Province, Northeast China, were examined for Cryptosporidium spp. and E. bieneusi using nested PCR. The overall infection rates for Cryptosporidium spp. and E. bieneusi were 6.38% (n = 27) and 7.09% (n = 30), respectively. The prevalence in different age groups ranged from 3.80% (95% confidence interval (CI) 1.01-6.59) to 8.36% (95% CI 4.83-11.90) for Cryptosporidium spp. and 5.97% (95% CI 2.52-9.43) to 7.94% (95% CI 4.49-11.40) for E. bieneusi. By analyzing the DNA sequences of the small subunit (SSU) rRNA gene, two Cryptosporidium species were detected in this study, namely C. parvum (n = 25) and C. ryanae (n = 2). The IIdA20G1 subtype was further identified by using the 60-kDa glycoprotein (gp60) gene of C. parvum. E. bieneusi was identified using three known sequences through the analysis of internal transcribed spacer (ITS) sequences: J (n = 23), I (n = 5), and BEB4 (n = 2), and all belonged to group 2. The results indicated that some of the Cryptosporidium species and E. bieneusi genotypes identified in Longjiang Wagyu cattle in the study areas might have zoonotic potential., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Objective: To observe the effect of early acupoint electrical stimulation on the decline of lower limbs muscle strength in patients with intensive care unit-acquired weakness (ICU-AW) caused by septic shock., Methods: A total of 58 patients with ICU-AW caused by septic shock were randomly divided into an observation group (28 cases, 1 case dropped off ) and a control group (30 cases, 2 cases dropped off ). Patients in both groups received routine basic treatment. In the observation group, acupoint electric stimulation therapy was added at Huantiao (GB 30), Futu (ST 32), Zusanli (ST 36), Xuanzhong (GB 39) and Taichong (LR 3). Unilateral point with electrodes were applied, the SDZ-Ⅱ electronic instrument (discontinuous wave, frequency in 2 Hz, strength in 5 mA) was connected and changed to the other side after 30 min of unilateral treatment.The treatment was given 2 times daily, continued for 7 d or until the medical research council (MRC) score being 54 points or more. The changes of lower limb muscle strength MRC score, modified Rankin scale (MRS) score, bilateral quadriceps thickness and gastrocnemius pinnate angle of both groups were observed before treatment and on discharge. The time of admission to ICU, time of hospitalization, mortality during hospitalization, and mortality 28 d after discharge were compared between the two groups. The MRS scores of the two groups were followed up 28 d after discharge., Results: The MRC scores of lower limb muscle strength in the two groups on discharge were higher than those before treatment ( P <0.05), and the MRS scores on discharge and 28 d after discharge in the two groups were lower than those before treatment ( P <0.05). The MRC scores of lower limb muscle strength on discharge in the observation group were higher than thoes in the control group ( P <0.05), and the MRS scores on discharge and 28 d after discharge in the observation group were lower than those in the control group ( P <0.05). On discharge, bilateral quadriceps thickness and gastrocnemius pinnate angle in the two groups were increased compared with those before treatment ( P <0.05), and thoese in the observation group was higher than the control group ( P <0.05). There was no significant difference between the two groups in the time of admission to ICU, time of hospitalization, mortality during hospitalization, and mortality 28 d after discharge ( P >0.05)., Conclusion: Early acupoint electrical stimulation can improve the lower extremity muscle decline in patients with ICU-AW caused by septic shock.

Although the effective transmission of the H5N1 virus from humans to humans has yet to be further observed, humans are at increased risk of a pandemic caused by H5N1. In order to fully evaluate the seroprevalence and risk factor of highly pathogenic avian influenza A (H5N1) virus infection among in mainland China, we performed a systematic review and meta-analysis. In this review, we searched literature on the seroprevalence of H5N1 infection among humans in mainland China from 1 January 1997 to 20 October 2018 in English and Chinese databases, including PubMed, Google scholar, Cochrane library, Clinical Trial, VIP, CNKI and WanFang database. We made a selection according to the title and the abstract of paper, and then, we excluded duplicated literature, and data incomplete literature according to the exclusion criteria we formulated. Finally, we extracted how many humans have H5N1 infection from the obtained studies to establish the seroprevalence of H5N1 infection among humans in mainland China. A total of 56 studies (including data of 35,159 humans) were compliant with our criteria. In China, the overall seroprevalence of H5N1 infection among humans was 2.45% (862/35,159), while the seroprevalence of H5N1 infection among humans from central China was 7.32% (213/2,911), higher than those in other regions of China. The seroprevalence of H5N1 infection was associated with test method, sampling time and demographic characteristics of humans. However, there was no significant difference in the effect of gender on the seroprevalence of H5N1 among humans in China. The purpose of this review was to better understand the real infection rate of H5N1 virus among humans and evaluate the potential risk factors for the zoonotic spread of H5N1 virus to humans. Sufficient epidemiological data are important to explore and understand the prevalent status of AIVs throughout the country and to disease control., (© 2020 Blackwell Verlag GmbH.)

Haemophilus parasuis, a gram-negative bacterium as an early commensal colonizer in the upper respiratory tract of weaning pigs (Sus scrofa), is one of the most important bacterial pathogens affecting pig populations. It is the causative agent of Glässer's disease, causing systemic infection and polyserositis, meningitis, and arthritis. H. parasuis infection can result in high mortality and morbidity with, the significant economic losses for pig producers. To estimate the overall disease prevalence of H. parasuis in pigs from China, we performed a meta-analysis using five bibliographical databases: PubMed, ScienceDirect, CNKI, Wanfang, and VIP Chinese Journal Databases. A total of 41 articles published between 2005 and 2019, fulfilled the final inclusion criteria. The overall prevalence of H. parasuis in pigs in China was 27.8 % with the highest prevalence between 2011 and 2015 (41.0 %). In terms of pig age, the point estimate of H. parasuis prevalence was higher in suckling piglets (29.2 %) compared with that for other pig ages. The prevalence in the serum subgroup (29.8 %) was higher than that in the nasal swab subgroup (12.5 %). The results of the present meta-analysis showed that H. parasuis infection was common in pig populations in China; therefore, effective control measures are necessary to reduce this threat to pig populations., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Hepatocellular carcinoma is one of the most common malignant tumors found all over the globe. Despite advances in surgery and chemotherapy, the five-year survival rate of patients with hepatocellular carcinoma is still low. It is known that the proliferation of hepatocellular carcinoma cells is closely related to the occurrence, development and prognosis of hepatocellular carcinoma. The present work investigates the expression of microRNA-489 (miR-489) in human hepatocellular carcinoma cells and its effect on the biological behavior of human hepatocellular carcinoma cells., Methods: The expression of miR-489 by fluorescence quantitative PCR detection in 30 patients with hepatoblastoma of liver cancer tissues and adjacent tissues was studied. Also, the determination of hepatoblastoma in four cell lines with different metastatic potential (HR8348, HCT116, HT29 and HEPG2) and the expression of miR-489 during miR-489 simulation process was studied. MTT assay, flow cytometry and Western blot analysis were performed to know the cell proliferation to detect the changes in cell cycle, apoptosis of cells, and SOX4 gene expression respectively., Results: RT-PCR results showed that the cells compared with pre-cancerous tissue, the expression level of miR-489 in hepatocellular carcinoma tissues than in adjacent tissue significantly decreased (P<0.05), and with liver cancer cell metastasis increased (P<0.05); analogue transfection constructed miR-489 overexpressing HEPG2 cell line by microRNA. MTT results showed that miR-489 can inhibit the proliferation of HEPG2 cells, the differences were statistically significant (P<0.05); flow cytometry results showed that miR-489 mimics was transfected into HEPG2 cells at 48 hours had no significant effect on cell cycle distribution (P > 0.05); but miR-489 expression could induce apoptosis, compared with the control group, the apoptosis of miR-489 mimics was significantly increased and the difference was statistically significant (P < 0.05)., Conclusion: In conclusion, miR-489 can significantly inhibit the occurrence and development of hepatocellular carcinoma cells. The mechanism may be down regulated by the expression of SOX4 and inhibit cell proliferation. Further this study showed that the tumor cells SOX4 gene as a regulatory factor target the genes of miR-489 in hepatocellular carcinoma., (© 2020 Wang B et al.)

Objective: To compare the therapeutic effect of transcutaneous electrical acupoint stimulation (TEAS) combined with conventional western medicine and western medicine alone on early enteral nutrition tolerance in patients with sepsis., Methods: Forty-nine patients with sepsis were randomly divided into an observation group (24 cases) and a control group (25 cases). The control group was treated with conventional western medicine, including limited fluid resuscitation, anti-infection and maintenance of hemodynamics, and enteral nutrition was provided when the hemodynamics became stable and the dosage of vasoactive drugs was reduced. The observation group, on the basis of the treatment of the control group, was treated with TEAS at Tianshu (ST 25), Shangjuxu (ST 37), Jiexi (ST 41), Diji (SP 8), Zusanli (ST 36), Zhongwan (CV 12) and Daheng (SP 15) when the enteral nutrition was provided (dilatational wave, 2 Hz/10 Hz, twice a day, 30 min each time), and the TEAS stopped when enteral nutrition was normal. The antral motility index (MI) was evaluated by ultrasonography on the first, third and fifth day into treatment in the two groups, and the duration from providing enteral nutrition to normal enteral nutrition was recorded. The acute gastrointestinal injury (AGI) classification, admission time of intensive care unit (ICU) and hospitalization time were compared., Results: The antral MI on the third and fifth day into treatment in the observation group was significantly higher than that in the control group ( P <0.05). The duration from providing enteral nutrition to normal enteral nutrition was (5.08±0.65) days in the observation group, which was shorter than (5.56±0.65) days in the control group ( P <0.05). The improvement of AGI classification after treatment in the observation group was significantly superior to the control group ( P <0.05). The hospitalization time in the observation group was shorter than that in the control group ( P <0.05)., Conclusion: TEAS combined with conventional western medicine treatment could significantly promote gastrointestinal peristalsis, improve the early intestinal nutrition tolerance in patients with sepsis, so as to shorten the time of reaching the standard of enteral nutrition and hospitalization time, which is better than conventional western medicine treatment alone.

Toxoplasmosis, one of the most common zoonoses worldwide, is caused by Toxoplasma gondii. T. gondii can infect almost all warm-blooded animals, including humans. Horses are an intermediate host of T. gondii, representing a potential risk for humans. To determine the T. gondii seroprevalence in horses worldwide, a global meta-analysis was conducted. A total of 35 publications were obtained by searching the PubMed, ScienceDirect, Chinese Web of knowledge (CNKI) and Wanfang databases. A total of 12,354 horses were assessed, of which 1580 were positive for T. gondii. The pooled overall seroprevalence of horses infected by T. gondii was 11.29%. No significant difference of T. gondii seroprevalence was observed between male and female horses. The seroprevalence of T. gondii in horses from different countries varied. Our findings suggest that toxoplasmosis is prevalent in horses worldwide. Therefore, it is necessary to implement continuous monitoring of the status of T. gondii seroprevalence in horses. Moreover, powerful regulatory measures should be implemented to prevent and control the spread of toxoplasmosis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The protistan pathogens Cryptosporidium and Enterocytozoon bieneusi can cause significant intestinal diseases in animals and humans. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium and E. bieneusi in ruminants in Northern China. In this study, the overall prevalence of Cryptosporidium and E. bieneusi was 19.3% (62/321) and 28.97% (93/321) in dairy calves and 1.10% (9/818) and 13.57% (111/818) in sika deer (Cervus nippon) in four provinces in Northern China, respectively. The prevalence of Cryptosporidium and E. bieneusi in different factor groups was various. Five Cryptosporidium species/genotypes were identified, of which C. parvum, C. ryanae, C. bovis, and C. andersoni were only found in dairy calves, and only Cryptosporidium deer genotype was found in sika deer. Moreover, J, I, and BEB4 ITS genotypes of E. bieneusi were found in dairy calves, and six known genotypes (JLD-III, JLD-IX, JLD-VII, EbpC, BEB6, and I) and ten novel genotypes (namely LND-I and JLD-XV to JLD-XXIII) were found in sika deer in this study. Cryptosporidium parvum and E. bieneusi genotype J were identified as the predominant species/genotypes in dairy calves, whereas the predominance of Cryptosporidium spp. and E. bieneusi in sika deer was Cryptosporidium deer genotype and BEB6, respectively. The present study reported the prevalence and genotypes of Cryptosporidium and E. bieneusi in dairy calves and sika deer in four provinces in northern China. The present findings also suggest that investigated dairy calves and sika deer may play an important role in the transmission of E. bieneusi and Cryptosporidium to humans and other animals, and also in an effort to better understand the epidemiology of these enteric pathogens in China.

Aims: Pseudomonas aeruginosa is one of the leading causes of opportunistic and hospital-acquired infections worldwide, which is frequently linked with clinical treatment difficulties. Ibuprofen, a widely used non-steroidal anti-inflammatory drug, has been previously reported to exert antimicrobial activity with the specific mechanism. We hypothesized that inhibition of P. aeruginosa with ibuprofen is involved in the quorum sensing (QS) systems., Main Methods: CFU was utilized to assessed the growth condition of P. aeruginosa. Crystal violent staining and acridine orange staining was used to evaluate the biofilm formation and adherence activity. The detection of QS virulence factors such as pyocyanin, elastase, protease, and rhamnolipids were applied to investigation the anti-QS activity of ibuprofen against P. aeruginosa. The production of 3-oxo-C 12 -HSL and C 4 -HSL was confirmed by liquid chromatography/mass spectrometry analysis. qRT-PCR was used to identify the QS-related gene expression. Furthermore, we explored the binding effects between ibuprofen and QS-associated proteins with molecular docking., Key Findings: Ibuprofen inhibits P. aeruginosa biofilm formation and adherence activity. And the inhibitory effects of ibuprofen on C 4 -HSL levels were concentration-dependent (p < 0.05), while it has no effect on 3-oxo-C 12 -HSL. Moreover, ibuprofen attenuates the production of virulence factors in P. aeruginosa (p < 0.05). In addition, the genes of QS system were decreased after the ibuprofen treatment (p < 0.05). Of note, ibuprofen was binding with LuxR, LasR, LasI, and RhlR at high binding scores., Significance: The antibiofilm and anti-QS activity of ibuprofen suggest that it can be a candidate drug for the treatment of clinical infections with P. aeruginosa., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Horse meat and milk are an important source of nutrition for many Chinese. Previous studies have shown a fairly high prevalence of Toxoplasma gondii infection in horses. However, there is no data concerning the molecular characterization of T. gondii in horses in China. The present study tested 231 cervical lymph node samples of horses from northern China (97 from Jilin, 54 from Liaoning, and 80 from Xinjiang) for the presence of the T. gondii B1 gene by seminested PCR. The B1-positive samples were genotyped at nine nuclear loci, that is, SAG1, alternative SAG2, SAG3, BTUB, c22-8, GRA6, c29-2, PK1, and an apicoplast locus, using PCR-restriction fragment length polymorphism technique. A total of 14 (6.06%) out of 231 samples were T. gondii positive. The overall prevalence of T. gondii in the horses was 5.15% (5/97) for Jilin Province, 5.55% (3/54) for Liaoning Province, and 7.50% (6/80) for Xinjiang Uygur Autonomous Region. Of the 14 positive samples, only 2 were successfully genotyped at all loci, 5 were successfully genotyped at 5-8 loci, and all typed samples belong to ToxoDB genotype no. 9. To our knowledge, this is the first molecular characterization of the T. gondii isolates from horses in China.

Atherosclerosis is a chronic inflammatory disease with lipid accumulation. Apolipoprotein C3 (APOC3), which is an important regulator of human lipid metabolism, is associated with multiple vascular mechanisms in atherosclerosis and proinflammatory responses. We have previously reported that the expression of inflammatory cytokine TNF-α is elevated in human endothelial cells (HUVECs) after APOC3 treatment. This study investigates the APOC3 signaling pathway involved in TNF-α-mediated expression of JAM-1 in HUVECs. Cultured HUVECs were exposed to APOC3 (50 μg/ml) for 16 h. Mechanistic studies were carried out by silencing TNF-α gene with lentiviral TNF-α-shRNA. Our study was based on the eight signaling pathway inhibitors to block the effect of APOC3 in HUVECs. The expression of JAM-1 was determined by qRT-PCR, Western blotting, and flow cytometry. IKK2 degradation and NF-κB p65 phosphorylation were determined by Western blotting. Our results showed that APOC3 significantly promoted the TNF-α-induced expression of JAM-1 in HUVECs. Inhibiting APOC3 reversed the TNF-α-induced overexpression of JAM-1. Moreover, APOC3 induced the expression of NF-κB p65 and degraded IκB. In conclusion, APOC3 promoted the expression of JAM-1 via the NF-κB, IKK2, and PI3K signaling pathway., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Toxoplasma gondii, the causative agent of toxoplasmosis, is carried by a range of intermediate hosts, including pigeons. Eating raw or undercooked pigeon meat, as is common in regions of China, can easily pass on a potentially fatal T. gondii infection. However, there are little data about the seroprevalence and genotypes of T. gondii infection in pigeons. In this study, a total of 963 pigeon blood samples were collected from Jilin Province, Liaoning Province, and Inner Mongolia Autonomous Region in China between August 2015 and December 2016. The overall seroprevalence of T. gondii was 10.80% (104/963), with 8.80% (41/466) in males and 12.68% (63/497) in females. The seroprevalence of T. gondii in different regions varied from 9.04% (17/188) in Jilin City to 13.16% (20/152) in Shenyang City, a relatively high prevalence. The seroprevalence of T. gondii in juvenile pigeons (6.38%, 18/282) was significantly lower than in adults (86/681, p < 0.05). These findings could provide useful data for the execution of effective control measures against T. gondii infection in pigeons and/or other hosts. More importantly, the findings also suggest that pigeons should not be eaten raw.

Background: Porcine epidemic diarrhea results from infection with porcine epidemic diarrhea virus (PEDV). It is an acute and highly contagious enteric disease in swine characterized by watery diarrhea and vomiting. Here, we performed a systematic review and meta-analysis in order to assess the prevalence of PEDV infection in pig populations from mainland China., Methods: We conducted a literature search on the prevalence of PEDV infection in pigs between Jan 1, 1988 and Aug 20, 2018 in English and Chinese databases, including PubMed, Google scholar, Cochrane library, Clinical Trials, VIP, CNKI and WanFang database. Selections were made based on the title and the abstract of paper, and duplicated literature was excluded along with other host studies, and data incomplete literature according to the exclusion criteria we formulated. Finally, we extracted the number of swine with PEDV infection from the obtained studies and provided information that permitted us to estimate the prevalence of PEDV infection in pigs in mainland China., Results: A total of 45 studies (including data from 15,990 pigs) met our evaluation criteria. In China, the overall estimated prevalence of PEDV infection in pigs was 44% (7113/15,990), while the estimated prevalence of PEDV infection in pigs from northern China was 37% (793/2136), lower than those in other regions of China. The prevalence of PEDV infection was associated with sampling season, category of pigs and clinical signs (diarrhea) in pigs. However, the prevalence of PEDV among pigs in China was not significantly associated with the effect of detected target genes, nor was it associated with date of study publication., Conclusion: Our findings suggest that PEDV infection is common among pigs in China. It is therefore necessary to carry out further research and monitor the prevalence of PEDV infection. Furthermore, powerful and effective regulatory measures should be taken in order to prevent the transmission and spread of PEDV among pig populations., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Toxoplasma gondii is an obligate intracellular apicomplexan parasite that can infect almost all homoiothermal animals, including domestic raccoon dogs (Nyctereutes procyonoides). However, related reports on T. gondii infection in domestic raccoon dogs were limited in China. Therefore, a serological investigation was undertaken to investigate the seroprevalence and risk factors for T. gondii infection in domestic raccoon dogs. A total of 962 serum samples were collected from Jilin, Liaoning, Heilongjiang and Hebei provinces, northern China between April 2016 and November 2017, and were detected by the indirect hemagglutination test (IHA). The seroprevalence of T. gondii infection was 7.28% in the overall surveyed raccoon dogs by IHA, which was different among the four provinces ranging from 6.54% to 7.57%. The seroprevalence of T. gondii infection in male and female raccoon dogs was 6.62% and 7.79%, respectively. Based on statistical analysis, age was regarded as an important risk factor for T. gondii infection in raccoon dogs in this study (P < 0.05). This study reported the seroprevalence and risk factors of T. gondii infection in domestic raccoon dogs in northern China, which provided essential data for prevention and control of T. gondii infection in raccoon dogs in Jilin province, Liaoning province, Heilongjiang province and Hebei province., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Toxoplasmosis is caused by the protozoon Toxoplasma gondii that can infect almost all warm-blooded hosts including horses and humans. Horse meat is one of the most important meat products for human consumption. However, data on seroprevalence of T. gondii in horses intended for human consumption in Northern China is scarce. Thus, a total of 614 serum samples were collected from Jilin Province and Inner Mongolia Autonomous Region (IMAR) between 2015 and 2017, and were detected the seroprevalence for T. gondii by indirect hemagglutination assay test. In the prevalence study, the overall seroprevalence of T. gondii in horses was 17.92% (110/614) by IHA test, at the cut- off 1:64. Horses collected from Changchun (13.58%, P = 0.02) has a significantly lower seroprevalence than those from Meihekou City (21.72%) and Tongliao City (22.81%). Moreover, a significant difference of T. gondii seroprevalence was found between horses ages at less than 12 months (22.75%) and ≥12 months or higher (16.42%, P = 0.002). To our knowledge, this is the first report of seroprevalence of T. gondii in horses in Jilin and IMAR, Northern China. Logistic regression suggested that age and geographical location of horses were identified as risk factors. These findings will provide the useful data for controlling and preventing toxoplasmosis in horses and/or other hosts in China., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Aims: To investigate the roles of Lats1/p-YAP1 pathway in TBI-induced neuronal apoptosis and neurological deficits in rats., Results: We found that Lats1 and YAP1 were expressed in cerebral cortex neurons of Sprague-Dawley rats, and the phosphorylation levels of Lats1 and YAP1 in injured regions were significantly increased after TBI. Furthermore, inhibition of Lats1 not only decreased the level of p-YAP1, but also attenuated neuronal apoptosis and neurological impairment., Conclusions: Our work demonstrates that inhibition of Lats1/p-YAP1 pathway mitigates neuronal apoptosis and neurological deficits in a rat model of TBI., (© 2018 John Wiley & Sons Ltd.)

Pig is the well-known intermediate host of T. gondii, a ubiquitous and obligate intracellular zoonotic pathogen. However, information about prevalence and genotypes of T. gondii infection in free-ranging pigs is scarce. Therefore a total of 186 hilar lymph nodes specimens were collected from free-ranging pigs from rural regions from Jilin (n = 119) and Liaoning (n = 67) provinces, northeastern China, and were investigated from T. gondii infection by semi-nested PCR of the B1 gene. Positive testing samples were genotyped by using polymerase chain reaction followed by restriction-fragment length polymorphism technology (PCR-RFLP) applied to 11 loci. The overall prevalence of T. gondii in investigated free-ranging pigs was 18.3% (34/186), with 16.4% (11/67) in Liaoning Province and 19.3% (23/119) in Jilin Province. Moreover, two genotypes, namely ToxoDB #9 and ToxoDB #10, were detected in investigated pigs. The present study showed a high T. gondii prevalence in free-ranging compared to farmed pigs as reported in previous studies by others, raising a major public health concern. It is essential to establish efficient strategies to prevent and control T. gondii infection in free-ranging pigs, other animals and humans in investigated regions., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Chlamydia is considered as one of the most widely prevalent zoonotic pathogens. It can spread from infected birds to human beings through direct or indirect contact with fecal shedding of Chlamydia. However, data concerning prevalence and genotypes of Chlamydia in pigeons are limited. In the present study, a total of 963 serum samples was collected from Jilin Province, Liaoning Province, and Inner Mongolia Autonomous Region (IMAR) in China between August 2015 and December 2016 and the seroprevalence for Chlamydia was analyzed by indirect hemagglutination assay test. The seroprevalence of Chlamydia was 20.4% (215/963) in total, at the cutoff 1:16, with the titers of 1:16 in 109, 1:64 in 49, 1:256 in 38, and 1:1024 in 18. Samples from all six administrative cities were detected Chlamydia-seropositive, ranging from 19.0% to 25.0%. Adult pigeons (23.5%) have a significant higher seroprevalence than juveniles (15.2%). Four PCR-positive samples represented Chlamydia psittaci genotype B. This is the first report of Chlamydia infection in pigeons in Liaoning Province and IMAR. The occurrence of C. psittaci genotype B in the droppings of pigeons suggests potential environmental contamination with C. psittaci and may raise a public health concern.

Aim: To evaluate the correlation between miRNA-375 and cell proliferation and apoptosis in glioma cancer cell., Methods: Collecting 30 cases of glioma cancer patients and 30 cases of cerebral infarction patients. The miRNA-375 and CTGF protein expressions were evaluated by ISH and IHC methods. In the cell experiment, the U87 cells were divided into 3 groups: NC group (the cells were treated with normal method); BL group (the cells were transfected with empty vector) and miRNA group (the cells were transfected with miRNA-375). The U87 cell proliferation and apoptosis rates and cell cycle of the different groups were measured by MTT and flow cytometry. The relative proteins (CTGF, EGFR, AKT, Erk and P21) expressions were measured by WB assay., Results: The miRNA-375 and CTGF expressions of glioma cancer tissues were significantly different compared with those of no-cancer tissues (p < 0.05, respectively). In the cell experiments, the cell proliferation of miRNA group was significantly decreased compared with that of NC group (p < 0.05); the cell apoptosis and G1 phase rate of miRNA group was significantly decreased compared with NC group (p < 0.05, respectively). Depending on the WB assay, the CTGF, EGFR, AKT, Erk and P21 proteins expressions of miRNA group were significantly different compared with proteins expressions of NC group (p < 0.05, respectively)., Conclusion: miRNA-375 over-expression suppresses glioma cancer cells development via CTGF-EGFR pathway (Fig. 3, Ref. 30).

Introduction: Previous studies have shown that hyperlordotic C1-C2 fusion was related to postoperative subaxial kyphosis. However, most of the patients in these studies were complicated with rheumatoid arthritis (RA). Moreover, no studies have specifically evaluated the relationship between C1-C2 fusion angle and cervical sagittal vertical axis (cSVA), T1 slope or cranial tilt (CRT) after posterior C1-C2 fusion. This study aimed to investigate the cervical sagittal alignment in non-RA patients following posterior C1-C2 fusion and the correlation between C1-C2 fusion angle and postoperative cervical sagittal alignment., Materials and Methods: From August 2004 to December 2015, twenty-eight consecutive patients with an average age of 39.2 years (range 6-70 years) who underwent posterior C1-C2 fusion from a single institution were enrolled. The mean follow-up period was 30.7 months (range 12-77 months). Angles of Oc-C1, C1-C2, C2-C3 and C2-C7, cSVA, T1 slope and CRT were measured in lateral cervical radiographs in neutral position before surgery and at the final follow-up., Results: C1-C2 angle significantly increased from 13.6° ± 12.4° to 22.0° ± 8.1° at the final follow-up (P < 0.001). A significant decrease was found both in Oc-C1 and C2-C7 angles from pre-operation to the final follow-up (P < 0.001 and P = 0.011, respectively). Moreover, cSVA and CRT dramatically increased from pre-operation to the final follow-up (P < 0.001). C1-C2 fusion angle was significantly associated with Oc-C1, C2-C7 angle, cSVA and CRT at the final follow-up. A significant correlation was also observed between postoperative change of C1-C2 angle and that of Oc-C1, C2-C7 angle, cSVA and CRT., Conclusions: Apart from decreased subaxial lordosis, posterior C1-C2 fusion in hyperextension may also lead to kyphotic change of atlanto-occipital alignment and increased tilting forward of the cervical spine. Therefore, intraoperative overreduction of C1-C2 angle and hyperlordotic C1-C2 fusion should be avoided to maintain the physiologic cervical sagittal alignment.

Objective: To investigate the epidemiology characteristics of crawfish related rhabdomyolysis (RM) in Nanjing, 2016., Methods: Outpatient and inpatient electronic medical system of 21 hospitals in Nanjing during 2016 were retrospectively searched, and all the patients diagnosed with RM were selected. The patients with none crayfish-related RM was excluded. The epidemiology characteristics were depicted. The geographic information system (GIS) was used to collect, manage and analyze the spatial data, to visualize it, to analyze the spatial distribution features of the disease, and to explore the cause of disease prediction. GeoDa 1.8 software was used to analyze the global and local spatial auto-correlation., Results: A total of 1 183 patients with crawfish related RM were initially screened, excluding 59 patients with RM caused by trauma, severe exercise, heat stroke, myositis, poisoning, drugs, and genetic diseases, and 1 124 patients were enrolled. The proportion of men was 36.48% (410/1 124) with an incidence of 12.54/100 thousands; while of women was 63.52% (714/1 124) with an incidence of 21.86/100 thousands. The median age at onset was 34 (28, 43) years. From July to August, the incidence of crawfish related RM was the highest, accounting for 96.53% of the total number of cases. The top four incidence areas were Pukou (41.54/100 thousands), Jianye (25.94/100 thousands), Qixia (25.73/100 thousands), Gulou (25.04/100 thousands), all of which were adjacent to the Yangtze River. Global spatial autocorrelation analysis showed: Moran I = 0.427, Z = 2.646, P = 0.003, suggesting that the crawfish related RM had positive spatial autocorrelation. The results showed that the spatial structure of crawfish related RM existed in Nanjing in 2016. Local spatial autocorrelation analysis showed that the "high-high" concentration areas were Pukou, Jianye and Liuhe. The incidences of above three areas which were the Nanjing section of the lower reaches of the Yangtze River flowed through the region and surrounding areas were higher than the overall incidence of Nanjing., Conclusions: The prevalence of crawfish related RM in Nanjing during 2016 had an obvious region-concentrated character and global spatial autocorrelation with the high prevalent regions mainly concentrated in the urban areas adjacent to the Yangtze River.

Objective: We screened and identified the differential expression of the methylation phenotype in the whole genome of colorectal laterally spreading tumor (LSTs)., Materials and Methods: 3 tissue samples of colorectal polypoid adenomas (PAs), 3 tissue samples of LSTs and 3 tissue samples of colon cancer were analyzed with a high-density gene chip, and about 450,000 methylation sites were detected covering approximately 95% of the CpG islands. The Delta Data screening was taken through a cluster analysis of methylation phenotype differential expression. 50 tissue samples each of PAs patients, LSTs patients, and colorectal cancer patients were selected. Methylation-specific PCR (MSP) was used to detect RASSF1A and WIF-1 methylation levels. He RT-PCR method was used to detect the relative mRNA expression levels for methylation expression identification., Results: The degree of LST methylation was higher than that of PAs, and 1234 genes were found to have a lower expression when compared to colorectal cancer samples. 764 genes had a higher expression when compared to colorectal cancer, and 559 genes lower expression when compared to PAs. The average methylation level of LSTs was higher than that of PAs, and lower than that of colorectal cancer. The chromosomal location was taken on these 1234 genes, which were higher than that of PAs, and lower than that of colorectal cancer; 518 genes were located on chromosome No. 2 (41.98%), 236 on No. 5 (19.12%), 357 on No. 8 (28.93%), and 123 on No. 10 (9.97%). According to clustering analysis, DNA differentially methylated sites were mainly on genes of cell adhesion molecules regulation, signaling pathways, energy transduction, cell cycle and apoptosis. The positive rate of RASSF1A and WIF-1 methylation in the tissues of LSTs patients were higher than that of PAs, and lower than that of colorectal cancer; differences were statistically significant (p<0.05). The relative expression levels of RASSF1A and WIF-1mRNA in the tissues of LSTs patients were lower than that of PAs, higher than that of colorectal cancer, and the difference was statistically significant (p<0.05)., Conclusions: The administration of high-density gene chip technology has a good application value to screen the differential expression of LSTs gene methylation phenotype. Results are consistent with the identification results.

Objective: To explore the role of miR-202 in multiple myeloma (MM) cells, and study the regulation of miR-202 on drug sensitivity of MM cells. Methods: miR-202 and BAFF mRNA levels were detected by real-time PCR. U266 cells were transfected with miR-202-mimics, miR-202-inhibitor, siBAFF and their negative controls. After above treatments, protein levels of Bcl-2 family and MAPK signaling pathway were detected by Western blot analysis, and the proliferation and apoptosis ability of MM cells were examined by WST-1, Annexin V-FLUOS assay, respectively. Results: The results showed that the expression of miR-202 in CD138+ MM cells (0.304±0.354) and U266 cells (0.052± 0.009) were lower than in normal controls (3.550 ± 1.126) ( P <0.001, P =0.009), whereas BAFF mRNA levels (5.700 ± 0.734, 9.576 ± 2.887) were higher than in normal controls (1.819 ± 0.853) ( P <0.001, P = 0.006). The proliferation ability of U266 cells transfected with miR-202 mimics was significantly inhibited than in control group [(56.04±0.021)% vs (18.89±0.32)%, P =0.002]. The result of Western blot showed that the expression of Bcl-2 decreased by about 24%, and the expression of Bax increased by about 124% in cells transfected with miR-202 mimics. The apoptosis rate in cells transfected with miR-202 mimics was significantly more than in control group [(49.60±4.89)% vs (26.20±1.28)%, P =0.029]. The apoptosis rate in miR-202 mimics combined with Bort group (51.23±5.41)% was higher as compared with Bort treatment alone (31.70±4.40)% or miR-202 mimics control combined with Bort group (27.94±4.04)%, ( P =0.047, P = 0.028), whereas the apoptosis rate in miR-202 mimics combined with Thal or Dex had no significant difference compared with miR-202 mimics control [(11.66±1.91)% vs (10.63±1.74)%, P =0.700; (16.35± 1.32)% vs (17.43 ± 1.95)%, P =0.400]. The inhibitory rate of cell growth in miR-202 mimics combined with Bort group was higher as compared with Bort treatment alone [(36.93±5.98)% vs (18.18±4.10)%, P = 0.029]. The expressions of p-JNK protein decreased in U266 cells transfected with miR-202 mimics and treated with Bort. Conclusion: miR-202 mimics combined with Bort could inhibit proliferation and induce apoptosis of U266 cells through negative regulating target gene BAFF, which further inhibited the JNK/SAPK signaling pathway.

Background: Hypernatremia is an uncommon but important electrolyte abnormality in intensive care unit patients. Sepsis is one of the most common causes of intensive care unit admission, but few studies about the role of hypernatremia in sepsis has been published yet. In this study, we aimed to explore the risk factors for developing hypernatremia in patients with sepsis, and the prognosis of patients with sepsis with or without hypernatremia was also assessed., Materials and Methods: In this retrospective cohort study of 51 septic intensive care unit patients at a single center, we examined the risk factors for the development of hypernatremia and the association of hypernatremia with clinical outcomes using univariate and multivariable analyses. Clinical outcomes such as mortality and hospital duration of patients with or without hypernatremia were also compared., Results: Acute Physiology and Chronic Health Evaluation II score (odds ratio = 1.15; 95% CI: 1.022-1.294) was found to be the only independent risk factor for hypernatremia in patients with sepsis. Moreover, patients developing hypernatremia during hospitalization showed significantly higher morbidity and mortality., Conclusions: Acute Physiology and Chronic Health Evaluation II score may be an independent risk factor for hypernatremia in patients with sepsis. Moreover, hypernatremia is strongly associated with worse outcome in sepsis., (Copyright © 2016 Southern Society for Clinical Investigation. Published by Elsevier Inc. All rights reserved.)

The mechanism underlying abnormally high transcription of the glial cell line-derived neurotrophic factor (GDNF) gene in glioma cells is not clear. In this study, to assess histone H3K9 acetylation levels in promoters I and II of the gdnf gene in normal human brain tissue, low- and high-grade glioma tissues, normal rat astrocytes, and rat C6 glioblastoma cells, we employed chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR), real-time PCR, and a pGL3 dual fluorescence reporter system. We also investigated the influence of treatment with curcumin, a histone acetyltransferase inhibitor, and trichostatin A (TSA), a deacetylase inhibitor, on promoter acetylation and activity and messenger RNA (mRNA) expression level of the gdnf gene in C6 cells. Compared to normal brain tissue, H3K9 acetylation in promoters I and II of the gdnf gene increased significantly in high-grade glioma tissues but not in low-grade glioma tissues. Moreover, H3K9 promoter acetylation level of the gdnf gene in C6 cells was also remarkably higher than in normal astrocytes. In C6 cells, curcumin markedly decreased promoter II acetylation and activity and GDNF mRNA expression. Conversely, all three measurements were significantly increased following TSA treatment. Our results suggest that histone H3K9 hyperacetylation in promoter II of the gdnf gene might be one of the reasons for its abnormal high transcription in glioma cells.