Expression profiles of host miRNAs and circRNAs and ceRNA network during Toxoplasma gondii lytic cycle.

Toxoplasma gondii is an opportunistic protozoan parasite that is highly prevalent in the human population and can lead to adverse health consequences in immunocompromised patients and pregnant women. Noncoding RNAs, such as microRNAs (miRNAs) and circular RNAs (circRNAs), play important regulatory roles in the pathogenesis of many infections. However, the differentially expressed (DE) miRNAs and circRNAs implicated in the host cell response during the lytic cycle of T. gondii are unknown. In this study, we profiled the expression of miRNAs and circRNAs in human foreskin fibroblasts (HFFs) at different time points after T. gondii infection using RNA sequencing (RNA-seq). We identified a total of 7, 7, 27, 45, 70, 148, 203, and 217 DEmiRNAs and 276, 355, 782, 1863, 1738, 6336, 1229, and 1680 DEcircRNAs at 1.5, 3, 6, 9, 12, 24, 36, and 48 h post infection (hpi), respectively. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses revealed that the DE transcripts were enriched in immune response, apoptosis, signal transduction, and metabolism-related pathways. These findings provide new insight into the involvement of miRNAs and circRNAs in the host response to T. gondii infection.

A method to protect the endometrium for microwave ablation treating types 1-3 uterine fibroids: a preliminary comparative study.

PURPOSE: To investigate the feasibility of percutaneous intrauterine instillation of chilled saline to protect the endometrium during microwave ablation (MWA) treating types 1-3 uterine fibroids. MATERIALS AND METHODS: Twenty-six patients with types 1-3 uterine fibroids were prospectively enrolled in an intrauterine saline instillation group (study group). The same number of patients with types 1-3 uterine fibroids who previously received MWA without endometrial protection were retrospectively included in a control group. Endometrial impairment was evaluated by hysteroscopy and magnetic resonance imaging (MRI). RESULTS: In the study group, hysteroscopy revealed an intact endometrium in 17 patients, congestion and reddening of the endometrium due to heat in 8 patients, and a burnt necrosis with a size < 1 cm on the functional layer of the endometrium in 1 patient. On MRI, in the study group, there were 17 (65.4%), 6 (23.1%), and 3 (11.5%) patients with grades 0, 1, and 2 endometrial impairment, respectively, but no grade 3 endometrial impairment. In the control group, there were 8 (30.8%), 8 (30.8%), 7 (26.9%), and 3 (11.5%) patients with grades 0, 1, 2, and 3 endometrial impairment, respectively. Endometrial impairment in the study group was significantly better than that in the control group (p = 0.006). One patient had puncture tunnel bleeding and no other complications occurred in the study group. CONCLUSION: Intraoperative percutaneous intrauterine instillation of chilled saline may be effective and safe in reducing the thermal damage to the endometrium caused by MWA for treating types 1-3 uterine fibroids.

Differential miRNA expression profiles in the bone marrow of Beagle dogs at different stages of Toxocara canis infection.

BACKGROUND: Toxocara canis is distributed worldwide, posing a serious threat to both human and dog health; however, the pathogenesis of T. canis infection in dogs remains unclear. In this study, the changes in microRNA (miRNA) expression profiles in the bone marrow of Beagle dogs were investigated by RNA-seq and bioinformatics analysis. RESULTS: Thirty-nine differentially expressed (DE) miRNAs (DEmiRNAs) were identified in this study. Among these, four DEmiRNAs were identified at 24 h post-infection (hpi) and all were up-regulated; eight DEmiRNAs were identified with two up-regulated miRNAs and six down-regulated miRNAs at 96 hpi; 27 DEmiRNAs were identified with 13 up-regulated miRNAs and 14 down-regulated miRNAs at 36 days post-infection (dpi). Among these DEmiRNAs, cfa-miR-193b participates in the immune response by regulating the target gene cd22 at 24 hpi. The novel_328 could participate in the inflammatory and immune responses through regulating the target genes tgfb1 and tespa1, enhancing the immune response of the host and inhibiting the infection of T. canis at 96 hpi. In addition, cfa-miR-331 and novel_129 were associated with immune response and self-protection mechanisms at 36 dpi. 20 pathways were significantly enriched by KEGG pathway analysis, most of which were related to inflammatory response, immune response and cell differentiation, such as Cell adhesion molecules (CAMs), ECM-receptor interaction and Focal adhesion. CONCLUSIONS: These findings suggested that miRNAs of Beagle dog bone marrow play important roles in the pathogenesis of T. canis infection in dogs and provided useful resources to better understand the interaction between T. canis and the hosts.

Specific Characteristic of Hyperplastic Callus in a Larger Cohort of Osteogenesis Imperfecta Type V.

Hyperplastic callus (HPC) is the most conspicuous features of osteogenesis imperfecta (OI) type V, of which accurate diagnosis and treatment are facing challenges. We investigate the clinical features, and impact factors of HPC in OI type V patients. In this retrospective single-center study, a total of 21 patients with type V OI confirmed by IFITM5 mutation were included. Radiological characteristics of bone were evaluated by X-rays, dual-energy X-ray absorptiometry, and computed tomography scan. Bone biopsy specimens were performed and stained by routine hematoxylin-eosin. The effects of bisphosphonates on HPC were investigated. Eleven patients (52.3%) had HPCs at 19 skeletal sites, 11 of which affected the femur. Three patients developed four (21.1%) HPCs after fractures, and 15 (78.9%) HPCs occurred in absence of bone fracture. The progress of HPCs was variable, of which most HPCs enlarged in the initial phase and remained stable, and only one HPC dwindled in size. One patient had a rapidly growing mass on the right humerus, and biopsy showed irregular trabeculae of woven bone and immature bone and cartilage in the loose and edematous collagenous network without signs of tumor. Bisphosphonates treatment had no significant effects on HPC of OI patients. HPC is the specific characteristic of OI type V patients, and its location, shape, size, and progression are variable, and the femur is the most frequently involved site. It is very important to make a diagnosis of HPC through detecting IFITM5 mutation and completing pathological diagnosis if necessary. The treatment of HPC is worth further exploration.

Prevalence and multilocus genotyping of Cryptosporidium spp. in cattle in Jiangxi Province, southeastern China.

Cryptosporidium is a genus of single-celled protozoa, infecting a wide range of animals and humans. Although Cryptosporidium infections of cattle have been reported in some provinces in China, there is no available information on the prevalence and predominant species of Cryptosporidium in cattle in Jiangxi province. To investigate the prevalence of Cryptosporidium in cattle in Jiangxi province of China, 556 fecal samples were collected from eight farms in four cities and the SSU rRNA locus of Cryptosporidium was amplified from the DNA of each fecal sample by PCR. The overall prevalence of Cryptosporidium was 12.8% (71/556) in cattle in Jiangxi province, with 24.3% (54/222) in Nanchang city, 7.8% (13/166) in Gao'an city, 3.7% (4/108) in Xinyu city, and 0.0% (0/60) in Ji'an city. The differences of the prevalence rates by region, breed, and age groups were statistically significant. All positive PCR products of Cryptosporidium were successfully sequenced and identified as three Cryptosporidium species, namely Cryptosporidium bovis (1/556, 0.18%), Cryptosporidium ryanae (7/556, 1.3%), and Cryptosporidium andersoni (63/556, 11.3%). Furthermore, 36 C. andersoni isolates were successfully classified into three MLST (multilocus sequence typing) subtypes based on four genetic loci (MS1, MS2, MS3, and MS16). The predominant MLST subtype was A4, A4, A4, A1 (n = 30). These findings not only revealed the prevalence and predominant species of Cryptosporidium in cattle in Jiangxi province, but also provided a baseline for studying the genetic structure of C. andersoni, offering a novel resource for better understanding of the epidemiology of Cryptosporidium infection in cattle in Jiangxi province, southeastern China.

First report of Eimeria and Entamoeba infection in alpacas (Vicugna pacos) in Shanxi Province, northern China.

Intestinal protozoa Eimeria and Entamoeba can infect many animal species including alpacas. However, data on the prevalence and pathogenicity of species of the two genera Eimeria and Entamoeba in alpacas in China is scarce. The current study was carried out to investigate the prevalence of Eimeria and Entamoeba in alpacas in two cities (Taiyuan and Xinzhou) in Shanxi Province, northern China, using PCR-based approaches. Eimeria spp. were only found in Taiyuan city, and the overall prevalence was 1.64%. All samples collected from male alpacas were PCR-negative for Eimeria. Four Eimeria-positive samples were tested positive as Eimeria lamae. The molecular prevalence of Entamoeba in alpacas was 18.03% (66/366), including 16.39% (50/305) in alpacas from Taiyuan city and 26.23% (16/61) from Xinzhou city, respectively. The Entamoeba prevalence in male alpacas (25.00%) was significantly higher than that in female alpacas (15.69%). Entamoeba bovis was the predominant species, and no Entamoeba histolytica infection was detected. Nine unique SSU rRNA gene sequences of Entamoeba were obtained which formed a new cluster. The results showed that sex and location might be the risk factors associated with prevalence of Eimeria spp., and sex might be the risk factor associated with prevalence of Entamoeba spp.. This is the first report of Entamoeba in alpacas worldwide. These findings expand our understanding of the prevalence and genetic diversity of Eimeria and Entamoeba in alpacas.

Effects of Bisphosphonates on Osteoporosis Induced by Duchenne Muscular Dystrophy: A Prospective Study.

OBJECTIVE: Duchenne muscular dystrophy (DMD) is a severe X-linked progressive neuromuscular disease that brings a significantly increased risk of osteoporosis and bone fractures. We prospectively evaluated the effects of oral and intravenous bisphosphonates on the bones of children with DMD. METHODS: This study included a total of 52 children with DMD. They were divided into zoledronic acid (ZOL), alendronate (ALN), and control groups according to bone mineral density (BMD) and history of fragility fractures. For 2 years, all patients took calcium, vitamin D, and calcitriol. Meanwhile, 17 patients received infusions of ZOL, and 18 patients received ALN. BMD, serum levels of alkaline phosphatase (ALP) and the cross-linked C-telopeptide of type I collagen (ß-CTX) were evaluated. RESULTS: After 24 months of treatment, the percentage changes in lumbar spine BMD were 23.2 ± 9.7% and 23.6 ± 8.8% in the ZOL and ALN groups (all P<.01 vs. baseline). The increases did not differ between the ZOL and ALN groups, but were significantly larger than those of the control group (P<.01). Serum ß-CTX and ALP levels, respectively, were decreased by 44.4 ± 18.0% and 31.9 ± 26.7% in the ZOL group and by 36.0 ± 20.3% and 25.8 ± 14.4% in the ALN group (all P<.01 vs. baseline). CONCLUSION: Zoledronic acid and alendronate had similar protective effects to increase bone mineral density and reduce bone resorption in children with DMD, which were superior to treatment of calcium, vitamin D, and calcitriol. ABBREVIATIONS: 25OHD = 25 hydroxyvitamin D; ALN = alendro-nate; ALP = alkaline phosphatase; ALT = alanine aminotransferase; BMD = bone mineral density; BP = bisphosphonate; Ca = calcium; ß-CTX = cross-linked C-telopeptide of type I collagen; DMD = Duchenne muscular dystrophy; FN = femoral neck; GC = glucocorticoid; LS = lumbar spine; ZOL = zoledronic acid.

Novel genotypes and multilocus genotypes of Enterocytozoon bieneusi in two wild rat species in China: potential for zoonotic transmission.

Enterocytozoon bieneusi is an opportunistic pathogen in immunodeficient patients. Although this pathogen has been reported in many domestic animals, few data are available about the occurrence of E. bieneusi in wild rats. In the current study, a total of 228 fecal samples from two wild rat species (Leopoldamys edwardsi and Berylmys bowersi) in China were examined by a nested PCR-based sequencing approach employing the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The overall prevalence of E. bieneusi in wild rats was 33.3% (76/228), with 35.1% (39/111) in L. edwardsi and 31.6% (37/117) in B. bowersi. Ten E. bieneusi genotypes (including four known and six novel genotypes) were identified, with the novel CQR-2 (n = 15) as the predominant genotype. Phylogenetic analysis indicated that ten genotypes in the present study belong to zoonotic group 1, which contains many genotypes in humans. Furthermore, multilocus sequence typing (MLST) analysis showed that 19 ITS-positive samples were successfully amplified at three microsatellites and one minisatellite, forming 18 multilocus genotypes (MLGs). This is the first report of E. bieneusi infection in the wild rats L. edwardsi and B. bowersi. Our findings suggest that wild rats could be a significant source of human infection, including contaminated food and water.

Complex and dynamic transcriptional changes allow the helminth Fasciola gigantica to adjust to its intermediate snail and definitive mammalian hosts.

BACKGROUND: The tropical liver fluke, Fasciola gigantica causes fasciolosis, an important disease of humans and livestock. We characterized dynamic transcriptional changes associated with the development of the parasite in its two hosts, the snail intermediate host and the mammalian definitive host. RESULTS: Differential gene transcription analysis revealed 7445 unigenes transcribed by all F. gigantica lifecycle stages, while the majority (n = 50,977) exhibited stage-specific expression. Miracidia that hatch from eggs are highly transcriptionally active, expressing a myriad of genes involved in pheromone activity and metallopeptidase activity, consistent with snail host finding and invasion. Clonal expansion of rediae within the snail correlates with increased expression of genes associated with transcription, translation and repair. All intra-snail stages (miracidia, rediae and cercariae) require abundant cathepsin L peptidases for migration and feeding and, as indicated by their annotation, express genes putatively involved in the manipulation of snail innate immune responses. Cercariae emerge from the snail, settle on vegetation and become encysted metacercariae that are infectious to mammals; these remain metabolically active, transcribing genes involved in regulation of metabolism, synthesis of nucleotides, pH and endopeptidase activity to assure their longevity and survival on pasture. Dramatic growth and development following infection of the mammalian host are associated with high gene transcription of cell motility pathways, and transport and catabolism pathways. The intra-mammalian stages temporally regulate key families of genes including the cathepsin L and B proteases and their trans-activating peptidases, the legumains, during intense feeding and migration through the intestine, liver and bile ducts. While 70% of the F. gigantica transcripts share homology with genes expressed by the temperate liver fluke Fasciola hepatica, gene expression profiles of the most abundantly expressed transcripts within the comparable lifecycle stages implies significant species-specific gene regulation. CONCLUSIONS: Transcriptional profiling of the F. gigantica lifecycle identified key metabolic, growth and developmental processes the parasite undergoes as it encounters vastly different environments within two very different hosts. Comparative analysis with F. hepatica provides insight into the similarities and differences of these parasites that diverged > 20 million years ago, crucial for the future development of novel control strategies against both species.

Molecular Detection and Genotyping of Toxoplasma gondii in Edward's Long-Tailed Rats (Leopoldamys edwardsi).

Toxoplasma gondii is an important zoonotic parasite infecting humans and various animals with a worldwide distribution. However, limited information is available on T. gondii infection in wild rats. The present study aimed to examine the prevalence and characterize the genotypes of T. gondii in wild rats in two regions of China. Brain tissues were collected from 111 Edward's long-tailed rats (Leopoldamys edwardsi) and 117 Bower's white-toothed rats (Berylmys bowersi) between November 2017 and January 2018. Genomic DNA was extracted and amplified by PCR targeting the T. gondii B1 gene. B1 gene-positive samples were genotyped at 10 genetic markers (SAG1, SAG2 [5', 3'] and [alternative], SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) using multilocus nested polymerase chain reaction/restriction fragment length polymorphism. Six (5.41%, 6/111) Edward's long-tailed rats from Chongqing Municipality were positive for T. gondii B1 gene, whereas no T. gondii infection was detected in Bower's white-toothed rats (n = 117) from Guangdong province. T. gondii prevalence in female and male rats was 1.77% (2/113) and 3.48 (4/115), respectively. Four of the six positive DNA samples were completely genotyped at 10 genetic loci and were identified as ToxoDB#20. The present study revealed the occurrence of T. gondii infection in Edward's long-tailed rats. These findings raised public health concerning about T. gondii infection in wild rats. These results provide reference data for understanding the distribution of T. gondii genotypes in wild rats in China.

First report of bovine viral diarrhea virus and Mycobacterium avium subspecies paratuberculosis infection in Tibetan sheep (Ovis aries) in Tibetan Plateau, China.

Bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP) are important pathogens, which cause serious disease in animals. However, information about BVDV and MAP infection in Tibetan sheep in China is limited. Two thousand one hundred and eighty-seven blood samples were collected from Tibetan sheep between April 2013 and March 2014 from the Tibetan Plateau and tested for BVDV and MAP antibodies using commercial ELISA kits. The overall seroprevalence of BVDV and MAP in Tibetan sheep was 36.7 and 11.29%, respectively. Furthermore, risk factor analysis indicated that the age of sheep was statistically significant associated with BVDV infection and the region was considered as the risk factor of MAP infection in sheep (P < 0.05), gender and season were not considered as risk factors. This is the first report of seroprevalence and risk factors associated with BVDV and MAP infection in Tibetan Sheep in China, which will provide baseline information for controlling BVDV and MAP infection in ruminants in the Tibetan Plateau, western China.

Molecular Detection and Genotyping of Toxoplasma gondii in Pigs for Human Consumption in Hunan Province, China.

Toxoplasma gondii is one of the most important sources of foodborne diseases. In this study, the molecular prevalence and genotypes of T. gondii were investigated in pigs in Hunan province, China. A total of 339 brain tissue samples of pigs were collected from April 2015 to December 2017 in Hunan province and were used to detect the T. gondii B1 gene. Of these, 34 (10%; 95% confidence interval: 8.7-12.6) samples were tested positive for the T. gondii B1 gene. Positive samples were genotyped at 10 genetic markers (SAG1, SAG2 [5' + 3' SAG2, alter. SAG2], SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) using polymerase chain reaction-restriction fragment length polymorphism technology. Moreover, one sample was identified as genotype ToxoDB#10 (Type I), and another sample was suspected to be unusual genotype ToxoDB#61 that has never been reported in China. This study showed that T. gondii is prevalent in pigs in Hunan province, posing a food safety threat to the public health in the investigated areas. Our result has implications for better understanding the genetic diversity of T. gondii infections in animals in China.

Toxoplasma gondii Infection in Farmed Wild Boars (Sus scrofa) in Three Cities of Northeast China.

The apicomplexan protozoan parasite Toxoplasma gondii is a widely distributed etiological agent of foodborne illness. This parasite can cause production losses in livestock and serious disease in humans through consumption of contaminated meat. Pig meat is the most likely source of human infection, and wild boars may play a role in the transmission of T. gondii by serving as a reservoir host. This study aimed to investigate the seroprevalence of antibodies to T. gondii among farmed wild boars in China. In an 11-month survey, a total of 882 serum samples were obtained from farmed wild boars from three cities (Jilin City, Siping City, and Baishan City) in Jilin province, Northeast China and were tested for antibodies specific for T. gondii. Using modified agglutination test and a cutoff titer of 1:25, the prevalence of T. gondii infection in the examined samples was 10.0% (88 of 882). The highest seroprevalence was observed in animals from Jilin city (15.3%, 43/281) and followed by Siping (11.4%, 30/263) and Baishan (4.4%, 15/338). Logistic regression analysis revealed a significant correlation between the investigated geographic region and T. gondii infection. In addition, prevalence was higher in females compared to males, and the highest prevalence was detected in piglets. These findings indicate that farmed wild boars may become a source of foodborne toxoplasmosis, posing a food safety threat to the public health in the investigated areas. Implementation of effective measures to control T. gondii infection in farmed wild boars in China may be warranted.

Seroprevalence and Risk Factors of Fascioliasis in Yaks, Bos grunniens, from Three Counties of Gansu Province, China.

The aim of this study was to determine the seroprevalence and risk factors of fascioliasis in yaks, Bos grunniens, from 3 counties of Gansu Province in China. A total of 1,584 serum samples, including 974 samples from white yaks from Tianzhu, 464 from black yaks from Maqu, and 146 from black yaks from Luqu County, were collected and analyzed using ELISA to detect IgG antibodies against Fasciola hepatica. The overall F. hepatica seroprevalence was 28.7% (454/1,584), with 29.2% in white yaks (284/974) and 27.9% in black yaks (170/610). The seroprevalence of F. hepatica in yaks from Tianzhu, Luqu, and Maqu was 29.2%, 22.6%, and 29.5%, respectively. Female yaks (30.9%) had higher F. hepatica seroprevalence than male yaks (23.4%). Also, F. hepatica seroprevalence varied by different age group from 24.1% to 33.8%. Further, the seroprevalence ranged from 21.8% to 39.1% over different seasons. Interestingly, the season and age of yaks were associated with F. hepatica infection in yaks in the investigated areas. These findings provided a basis for further studies on this disease in yaks from 3 counties of Gansu Province in northwestern China, which may ultimately support the development of effective control strategies of fascioliasis in these areas.

Prevalence and risk factors of Cryptosporidium infection in farmed pigs in Zhejiang, Guangdong, and Yunnan provinces, China.

Cryptosporidiosis is caused by species of Cryptosporidium protozoa that can infect a wide range of host animals worldwide. However, data regarding Cryptosporidium infection in farmed pigs in subtropical areas in China is limited. Therefore, a total of 396 fecal samples were obtained from farmed pigs from Zhejiang (n = 124), Guangdong (n = 72), and Yunnan (n = 200) provinces, China, and were tested by PCR amplification of the small subunit (SSU) rRNA gene. The overall prevalence of Cryptosporidium in pigs was 17.68% (70/396), with 20.11% (36/179) in male pigs and 15.67% (34/217) in female pigs. Additionally, Cryptosporidium prevalence was 8.33% (6/72) in Guangdong province, 14.52% (18/124) in Zhejiang province, and 23.00% (46/200) in Yunnan province. A DNA sequence analysis of the SSU rRNA gene revealed that all of the isolates represented C. scrofarum. This is the first report of C. scrofarum infection in pigs in Guangdong and Yunnan provinces in subtropical areas of China. The results of the present study provide foundation data for control and prevention of Cryptosporidium infection in pigs in the study areas in China.

Prevalence, Risk Factors and Multilocus Genotyping of Giardia intestinalis in Dairy Cattle, Northwest China.

Giardia intestinalis is a cosmopolitan protozoan parasite that can infect a range of animals, including dairy cattle. As information regarding the prevalence and genotyping of G. intestinalis infection in dairy cattle in northwestern China is limited, 2,945 feces samples from 1,224 dairy cattle in Gansu Province and from 1,614 in Ningxia Hui Autonomous Region (NXHAR) were examined between December 2012 and March 2014. The overall prevalence of G. intestinalis was 3.63% (107/2,945), with 2.63% and 4.38% in Gansu and NXHAR, respectively. Logistic regression analysis showed region, age and season to be significant risk factors for G. intestinalis infection. Assemblage analysis identified 106 assemblage E and one assemblage A at the triose phosphate isomerase (tpi) locus in this study. Intravariations were also detected at tpi, glutamate dehydrogenase (gdh) and beta giardin (bg) loci within assemblage E, showing seven, three, and five new subtypes, respectively. Moreover, 13 new multilocus genotypes (E20-E32) were observed in assemblage E. Effective strategies and measures should be taken to prevent and control giardiasis in Gansu and NXHAR.

Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces.

Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.

[Recombinase Polymerase Amplification and its Applications in Parasite Detection].

Recombinase polymerase amplification (RPA) is a recently -developed isothermal nucleic-acid-amplification technology that is based on the nucleic acid replication mechanism in T4 bacteriophage. With this technique, nucleic-acid templates can be amplified to measurable levels within 20 min at 37-42 °C. The. RPA process has high sensitivity and specificity, and is simple to operate, thus nucleic acids can be detected rapidly in non-laboratory conditions. Since its development in 2006, the RPA technique has been applied in agriculture, food safety, medicine, transgene detection, etc. In this review, we will give an overview on the research progress of RPA and its application in parasite detection.

Proteomic change in the upper lobe of the left lung of Beagle dogs at the lung migration stage of Toxocara canis infection.

BACKGROUND: Toxocara canis is considered one of the most neglected parasitic zoonoses and threatens the health of millions of people worldwide with a predilection for pediatric and adolescent populations in impoverished communities. Exploring the invasion and developmental mechanisms associated with T. canis infection in its definitive canine hosts will help to better control zoonotic toxocariasis. METHODS: Proteomic changes in samples from the upper lobe of the left lung of Beagle puppies were systematically analyzed by quantitative proteomic technology of data-independent acquisition (DIA) at 96 h post-infection (hpi) with T. canis. Proteins with P-values < 0.05 and fold change > 1.5 or < 0.67 were considered proteins with differential abundance (PDAs). RESULTS: A total of 28 downregulated PDAs and 407 upregulated PDAs were identified at 96 hpi, including RhoC, TM4SFs and LPCAT1, which could be associated with the maintenance and repair of lung homeostasis. GO annotation and KEGG pathway enrichment analyses of all identified proteins and PDAs revealed that many lung proteins have correlation to signal transduction, lipid metabolism and immune system. CONCLUSIONS: The present study revealed lung proteomic alterations in Beagle dogs at the lung migration stage of T. canis infection and identified many PDAs of Beagle dog lung, which may play important roles in the pathogenesis of toxocariasis, warranting further experimental validation.

The Toxoplasma monocarboxylate transporters are involved in the metabolism within the apicoplast and are linked to parasite survival.

The apicoplast is a four-membrane plastid found in the apicomplexans, which harbors biosynthesis and organelle housekeeping activities in the matrix. However, the mechanism driving the flux of metabolites, in and out, remains unknown. Here, we used TurboID and genome engineering to identify apicoplast transporters in Toxoplasma gondii. Among the many novel transporters, we show that one pair of apicomplexan monocarboxylate transporters (AMTs) appears to have evolved from a putative host cell that engulfed a red alga. Protein depletion showed that AMT1 and AMT2 are critical for parasite growth. Metabolite analyses supported the notion that AMT1 and AMT2 are associated with biosynthesis of isoprenoids and fatty acids. However, stronger phenotypic defects were observed for AMT2, including in the inability to establish T. gondii parasite virulence in mice. This study clarifies, significantly, the mystery of apicoplast transporter composition and reveals the importance of the pair of AMTs in maintaining the apicoplast activity in apicomplexans.