Search

Abstract Background Ischemic stroke (IS) is a common and serious neurological condition that is highly fatal but so far no early diagnostic markers are available. Myocardial infarction-associated transcript (MIAT) is a long non-coding RNA (lncRNA) that could lead to IS by inducing autophagy and apoptosis in neuronal cells. However, there has been no report on the link between susceptibility to IS and the single-nucleotide polymorphisms (SNPs) of MIAT. This study aimed to investigate the association between MIAT gene polymorphisms and IS risk. Methods A total of 320 IS patients and 310 age-, sex- and race-matched controls were included in this study. Four polymorphisms (rs2157598, rs5761664, rs1894720, and rs9625066) were genotyped by using SNPscan technique. Results Among the 4 polymorphisms of MIAT, only rs9625066 was associated with IS risk (CA vs. CC: adjusted OR = 0.55, 95% CI, 0.37–0.85, P = 0.006; AA vs. CC: adjusted OR = 0.39, 95% CI, 0.16–0.94, P = 0.036; (AA + CA vs. CC: adjusted OR = 0.53, 95% CI, 0.35–0.80, P = 0.002; A vs. C adjusted OR = 0.59, 95% CI, 0.42–0.82, P = 0.002). Haplotype analysis showed a 1.32-fold increase (95% CI, 1.05–1.67, P = 0.017) in IS risk for rs2157598-rs5761664-rs1894720-rs9625066 (A-C-G-C). Logistic regression analysis identified some independent impact factors for IS including rs9625066 AA/AC, TC, TG, HDL-C (P

Abstract Background The miR-208 gene is one of the microRNAs now under active studies, and has been found to play significant roles in an array of cardiovascular diseases. Nevertheless, until now, no studies have examined the relationship between the susceptibility to ischemic stroke (IS) and genetic variations in miR-208. This study explored the association between the miR-208 polymorphisms (rs178642, rs8022522, and rs12894524) and the risk of IS. Methods A total of 205 cases of IS and 211 control subjects were included. The SNPscans genotyping test was employed to determine the genotypes of the three polymorphisms. Results Significant correlation was observed between rs8022522 polymorphism and risk of IS on the basis of analyses of genotypes, models and alleles (GA vs. GG: adjusted OR = 2.159, 95% CI: 1.052–4.430, P = 0. 036; AA vs. GG: adjusted OR = 5.154, 95% CI: 1.123–23.660, P = 0.035; dominant model: adjusted OR = 1.746, 95% CI, 1.075–2.838, P = 0.025; G vs. A: adjusted OR = 2.451, 95% CI: 1.374–4.370, P = 0.002). Conclusions The rs8022522 polymorphism of the miR-208 gene is significantly associated with an elevated risk of ischemic stroke in Chinese.

Sugarcane, a vital cash crop, contributes significantly to the world’s sugar supply and raw materials for biofuel production, playing a significant role in the global sugar industry. However, sustainable productivity is severely hampered by biotic and abiotic stressors. Genetic engineering has been used to transfer useful genes into sugarcane plants to improve desirable traits and has emerged as a basic and applied research method to maintain growth and productivity under different adverse environmental conditions. However, the use of transgenic approaches remains contentious and requires rigorous experimental methods to address biosafety challenges. Clustered regularly interspaced short palindromic repeat (CRISPR) mediated genome editing technology is growing rapidly and may revolutionize sugarcane production. This review aims to explore innovative genetic engineering techniques and their successful application in developing sugarcane cultivars with enhanced resistance to biotic and abiotic stresses to produce superior sugarcane cultivars.

Abstract Background The impact of host skin microbiome on horizontal transmission of tick-borne pathogens , and of pathogen associated transstadial and transovarial changes in tick microbiome are largely unknown, but are important to control increasingly emerging tick-borne diseases worldwide. Methods Focusing on a rickettsiosis pathogen, Rickettsia raoultii, we used R. raoultii-positive and R. raoultii-negative Dermacentor spp. tick colonies to study the involvement of skin microbiota in cutaneous infection with rickettsiae in laboratory mice, and the function of the tick microbiome on maintenance of rickettsiae through all tick developmental stages (eggs, larvae, nymphs, adults) over two generations. Results We observed changes in the skin bacteria community, such as Chlamydia, not only associated with rickettsial colonization but also with tick feeding on skin. The diversity of skin microbiome differed between paired tick-bitten and un-bitten sites. For vertical transmission, significant differences in the tick microbiota between pathogenic rickettsia-positive and -negative tick chorts was observed across all developmental stages at least over two generations, which appeared to be a common pattern not only for R. raoultii but also for another pathogenic species, Candidatus Rickettsia tarasevichiae. More importantly, bacterial differences were complemented by functional shifts primed for genetic information processing during blood feeding. Specifically, the differences in tick microbiome gene repertoire between pathogenic Rickettsia-positive and -negative progenies were enriched in pathways associated with metabolism and hormone signals during vertical transmission. Conclusions We demonstrate that host skin microbiome might be a new factor determining the transmission of rickettsial pathogens through ticks. While pathogenic rickettsiae infect vertebrate hosts during blood-feeding by the tick, they may also manipulate the maturation of the tick through changing the functional potential of its microbiota over the tick’s life stages. The findings here might spur the development of new-generation control methods for ticks and tick-borne pathogens. Video Abstract

Abstract Typically, airway remodeling caused by migration and proliferation of airway smooth muscle cells (ASMCs) plays a crucial role in the pathophysiological characteristics of asthma development. Cystatin 1 (CST1), a protein‐coding gene referred to as Cystatin SN, is highly expressed in asthma patients. However, the role of CST1 and related molecular mechanisms in the development of asthma remains to be explored. This study aims to investigate the role of CST1 in asthma progression and present related molecular mechanisms. To explore these aspects, human ASMCs with platelet‐derived growth factor BB (PDGF‐BB) are initially stimulated and applied as a cellular model of asthma. Further, CST1 is knocked down with small interfering ribose nucleic acid (siRNA) overexpressed with plasmids. Then, 5‐ethynyl‐2′‐deoxyuridine (EdU) and Cell Count Kit (CCK)‐8 assays are applied to assess the cell proliferation rates. Further, Transwell and Western blot analyses for migration of cells and expression of MMP1 and MMP9 proteins are assessed, respectively. Under PDGF‐BB stimulation, human ASMCs showed an increased CST1 expression, enhanced proliferation, and migration abilities, as well as up‐regulated PI3K/AKT signaling pathway. Further, knockdown or overexpression of CST1 presented the declined or enhanced proliferation, migration, and up‐regulation of the PI3K/AKT signaling pathway of human ASMCs. Inhibiting PI3K/AKT signaling pathway displayed the reduced migration and proliferation of human ASMCs. In summary, these findings indicated that CST1 played an essential role in the progression of asthma by activating the PI3K/AKT signaling pathway and promoting the migration and proliferation abilities of human ASMCs treated with PDGF‐BB.

BackgroundIschemic stroke (IS) represents a major cause of morbidity and mortality across the globe. The aberrant expression of miR-365 has been found to be implicated in a wide array of human diseases, including atherosclerosis and cancer. Studies on single-nucleotide polymorphisms (SNPs) in miRNA genes can help gain insight into the susceptibility to the condition. This study aimed to examine the relationship between miR-365 SNPs and the risk of IS.MethodsThe study recruited 215 IS patients and 220 controls. The SNPscans genotyping was employed to genotype three polymorphic loci (rs121224, rs30230, and rs178553) of miR-365. The relative expression of miR-365 in peripheral blood mononuclear cells of the patients and controls was determined by using real-time quantitative PCR.ResultsThe miR-365 rs30230 polymorphism exhibited a significant association with the risk of developing IS (TC vs. CC: adjusted OR = 0.55, 95% CI: 0.33-0.92, P = 0.022; TT vs. CC: adjusted OR = 0.34, 95% CI: 0.14–0.85, P = 0.021; TC +TT vs. CC: adjusted OR = 0.51, 95% CI: 0.31–0.83, P = 0.007; T vs. C: adjusted OR = 0.57, 95% CI: 0.39–0.83, P = 0.004). Haplotype analysis revealed that the C-T-G haplotype was associated with a decreased risk of IS (OR = 0.68, 95% CI: 0.46–1.00, P = 0.047). Furthermore, miR-365 expression was significantly higher in IS patients than in controls (P < 0.001). Interestingly, patients with rs30230 TC or TT genotypes had lower miR-365 levels compared to their counterparts with CC genotypes (P < 0.001).ConclusionsThe miR-365 rs30230 polymorphism might bear an association with IS susceptibility in the Chinese population, and the rs30230 TC/TT genotype might be a protective factor against IS.

The present study was aimed at assessing more accurately the seismic vulnerability and performance of highway bridges in an earthquake-damaged area. Based on mathematical statistics (data collection) and a probability model, (i) survey data for 2134 bridges in 22 highway sections (47 sub-sections) affected by the Wenchuan earthquake on May 12, 2008 were obtained and analysed, (ii) Gaussian and exponential regression models were developed, (iii) the empirical seismic-damage vulnerability function, plane, surface and curve model of the 22 highway sections with sample number and failure ratio were established, and (iv) a regression parameter matrix was constructed considering the empirical seismic-damage data. Using the method of probability exceedance, the bridge vulnerability curve models considering the seismic-damage exceedance probability of each section were analysed, and a parameter matrix model (mean seismic-damage index) was proposed to evaluate and predict the seismic vulnerability of regional bridges. Based on creating the actual seismic-damage sample database, vulnerability point-cloud models of the 22 highway bridges were constructed, and the vulnerability probability matrix, function, curve and point-cloud model of each highway section can be used to evaluate and estimate the vulnerability of regional bridges in the future.

Abstract NASCION‐type lithium (Li) conductors provide a great chance to break the challenges of solid‐state lithium batteries (SSLBs) emphasizing superior safety and high energy density. Nonetheless, their practical employment has been hampered by the poor interfacial compatibility. Herein, we successfully block interfacial side reactions by in situ constructing a LiF‐enrich solid electrolyte interphase (SEI) layer between Li metal and LAGP (Li1.5Al0.5Ge1.5(PO4)3) through dropping trace fluoroacetonitrile‐based all‐fluorinated electrolyte. Noted that the formed high Young's modulus but fast‐kinetics LiF‐rich SEI layer successfully suppresses growth of Li dendrite, further tailoring the superior interfacial chemistry. Consequently, such robust SEI upgrades critical current density of LAGP to a record‐high value of >1.5 mA cm−2. Furthermore, a hybride full cells assembled with the commercial‐level cathode deliver prominently cycling lifespan (>250 cycles) and outstanding rate performance. The present SEI engineering strategy enables a huge leap toward the industrialized deployments of SSLBs.

IntroductionTicks are the most important obligate blood-feeding vectors of human pathogens. With the advance of high-throughput sequencing, more and more bacterial community and virome in tick has been reported, which seems to pose a great threat to people.MethodsA total of 14 skin specimens collected from tick-bite patients with mild to severe symptoms were analyzed through meta-transcriptomic sequencings.ResultsFour bacteria genera were both detected in the skins and ticks, including Pseudomonas, Acinetobacter, Corynebacterium and Propionibacterium, and three tick-associated viruses, Jingmen tick virus (JMTV), Bole tick virus 4 (BLTV4) and Deer tick mononegavirales-like virus (DTMV) were identified in the skin samples. Except of known pathogens such as pathogenic rickettsia, Coxiella burnetii and JMTV, we suggest Roseomonas cervicalis and BLTV4 as potential new agents amplified in the skins and then disseminated into the blood. As early as 1 day after a tick-bite, these pathogens can transmit to skins and at most four ones can co-infect in skins.DiscussionAdvances in sequencing technologies have revealed that the diversity of tick microbiome and virome goes far beyond our previous understanding. This report not only identifies three new potential pathogens in humans but also shows that the skin barrier is vital in preventing horizontal transmissions of tick-associated bacteria or virus communities to the host. It is the first research on patients’ skin infectome after a tick bite and demonstrates that more attention should be paid to the cutaneous response to prevent tick-borne illness.

Ticks are obligate blood-sucking vectors for multiple zoonotic diseases. In this study, tick samples were collected from Yunnan Province, China, which is well-known as the “Global Biodiversity Hotspot” in the world. This study aimed to clarify the microbial populations, including pathogens, associated with ticks and to identify the diversity of tick-borne microbiota in this region. The 16S rRNA full-length sequencing from pooled tick DNA samples and PCR amplification of pathogenic genera from individual samples were performed to understand tick-associated microbiota in this region. A total of 191 adult ticks of 5 tick species were included and revealed 11 phyla and 126 genera bacteria, including pathogenic Anaplasma , Ehrlichia , Candidatus Neoehrlichia, Rickettsia , Borrelia , and Babesia . Further identification suggested that Rickettsia sp. YN01 was a variant strain of Rickettsia spp. IG-1, but Rickettsia sp. YN02 and Rickettsia sp. YN03, were potentially two new SFGR species. This study revealed the complexity of ecological interactions between host and microbe and provided insight for the biological control of ticks. A high microbial diversity in ticks from Yunnan was identified, and more investigation should be undertaken to elucidate the pathogenicity in the area.

The Jiaodong Peninsula in Shandong Province, China is the world’s third-largest gold metallogenic area, with cumulative proven gold resources exceeding 5000 t. Over the past few years, breakthroughs have been made in deep prospecting at a depth of 500–2000 m, particularly in the Sanshandao area where a huge deep gold orebody was identified. Based on previous studies and the latest prospecting progress achieved by the project team of this study, the following results are summarized. (1) 3D geological modeling results based on deep drilling core data reveal that the Sanshandao gold orefield, which was previously considered to consist of several independent deposits, is a supergiant deposit with gold resources of more than 1200 t (including 470 t under the sea area). The length of the major orebody is nearly 8 km, with a greatest depth of 2312 m below sea level and a maximum length of more than 3 km along their dip direction. (2) Thick gold orebodies in the Sanshandao gold deposit mainly occur in the specific sections of the ore-controlling fault where the fault plane changes from steeply to gently inclined, forming a stepped metallogenic model from shallow to deep level. The reason for this strong structural control on mineralization forms is that when ore-forming fluids migrated along faults, the pressure of fluids greatly fluctuated in fault sections where the fault dip angle changed. Since the solubility of gold in the ore-forming fluid is sensitive to fluid pressure, these sections along the fault plane serve as the target areas for deep prospecting. (3) Thermal uplifting-extensional structures provide thermodynamic conditions, migration pathways, and deposition spaces for gold mineralization. Meanwhile, the changes in mantle properties induced the transformation of the geochemical properties of the lower crust and magmatic rocks. This further led to the reactivation of ore-forming elements, which provided rich materials for gold mineralization. (4) It can be concluded from previous research results that the gold mineralization in the Jiaodong gold deposits occurred at about 120 Ma, which was superimposed by nonferrous metals mineralization at 118–111 Ma. The fluids were dominated by primary mantle water or magmatic water. Metamorphic water occurred in the early stage of the gold mineralization, while the fluid composition was dominated by meteoric water in the late stage. The S, Pb, and Sr isotopic compositions of the ores are similar to those of ore-hosting rocks, indicating that the ore-forming materials mainly derive from crustal materials, with the minor addition of mantle-derived materials. The gold deposits in the Jiaodong Peninsula were formed in an extensional tectonic environment during the transformation of the physical and chemical properties of the lithospheric mantle, which is different from typical orogenic gold deposits. Thus, it is proposed that they are named “Jiaodong-type” gold deposits.©2021 China Geology Editorial Office.

BackgroundEstimating the growth of pulmonary sub-solid nodules (SSNs) is crucial to the successful management of them during follow-up periods. The purpose of this study is to (1) investigate the measurement sensitivity of diameter, volume, and mass of SSNs for identifying growth and (2) seek to establish a deep learning-based model to predict the growth of SSNs.MethodsA total of 2,523 patients underwent at least 2-year examination records retrospectively collected with sub-solid nodules. A total of 2,358 patients with 3,120 SSNs from the NLST dataset were randomly divided into training and validation sets. Patients from the Yibicom Health Management Center and Guangdong Provincial People’s Hospital were collected as an external test set (165 patients with 213 SSN). Trained models based on LUNA16 and Lndb19 datasets were employed to automatically obtain the diameter, volume, and mass of SSNs. Then, the increase rate in measurements between cancer and non-cancer groups was studied to evaluate the most appropriate way to identify growth-associated lung cancer. Further, according to the selected measurement, all SSNs were classified into two groups: growth and non-growth. Based on the data, the deep learning-based model (SiamModel) and radiomics model were developed and verified.ResultsThe double time of diameter, volume, and mass were 711 vs. 963 days (P = 0.20), 552 vs. 621 days (P = 0.04) and 488 vs. 623 days (P< 0.001) in the cancer and non-cancer groups, respectively. Our proposed SiamModel performed better than the radiomics model in both the NLST validation set and external test set, with an AUC of 0.858 (95% CI 0.786–0.921) and 0.760 (95% CI 0.646–0.857) in the validation set and 0.862 (95% CI 0.789–0.927) and 0.681 (95% CI 0.506–0.841) in the external test set, respectively. Furthermore, our SiamModel could use the data from first-time CT to predict the growth of SSNs, with an AUC of 0.855 (95% CI 0.793–0.908) in the NLST validation set and 0.821 (95% CI 0.725–0.904) in the external test set.ConclusionMass increase rate can reflect more sensitively the growth of SSNs associated with lung cancer than diameter and volume increase rates. A deep learning-based model has a great potential to predict the growth of SSNs.

Dongping Lake area, located in the lower reaches of Yellow River, is an ideal place to study the changes of modern river and lake sedimentary environment. The sediment samples of Dawen River, Yellow River, and Dongping Lake were collected, and the major elements, trace elements and organic matter geochemical composition of the samples were analyzed. Cluster analysis, characteristic element ratio method and graphic method were used to explore the geochemical characteristics of sediments and their environmental implication. The results show that the contents of SiO2, Na2O, TiO2 and Zr in sediments of Dawen River and Yellow River are relatively high, and the contents of iron and manganese oxides, organic matter, CaO, P2O5 and Sr in lake sediments are relatively high. That reveals the differences of sedimentary environments between the rivers and the lake. The contents of Sr and Zr in Dawen River are affected by the rapid migration of clastic materials in the upstream carbonate source area during the flood season; the δCe, ΣREE and REE’s ratios in the sediments of the Yellow River reflect the influence of the Loess source; and the distribution of elements changes along the flow direction during the flood season. The characteristics of pH, element composition and LREE & HREE fractionation of the lake sediments indicate that the sediment source is complex, and the lake environment is affected by the flood season. The study shows that the geochemical content and its variation characteristics of sediments effectively reveal the sedimentary environment, material composition and characteristics of flood season of rivers and the lake in the study area.

The macroseismic intensity scale in different regions is extensively assessed in the vulnerability assessment of building structures and is rarely used to quantify the damage characteristics of group bridges. To more accurately examine the differences in the seismic vulnerability of typical bridges in an earthquake-damaged region under multiple intensity scales, this study analyses the vulnerability of 1516 girder bridges and 612 arch bridges affected by the Wenchuan earthquake, which occurred on May 12, 2008, in China, by using the European Macroseismic scale, Medvedev, Sponheuer, and Karnik scale, and the Chinese Seismic Intensity scale based on the principle of nonlinear regression and probability models. The vulnerability matrices, functions, and curves of the update models to compare girder bridges and arch bridges were obtained based on the number of samples and their failure ratio. When combined with the new cumulative exceeding probability model, curves of the exceeding probability model of girder bridges and arch bridges under the different intensity scales were obtained and compared. A parameter matrix model is proposed to evaluate and predict the seismic vulnerability of regional bridges. Combined with empirical data on the seismic damage caused to the girder bridges and arch bridges in the earthquake, their vulnerability matrices, curves, and point cloud models were obtained under a multi-intensity scale of assessment for a comparative analysis. The conclusions of this study can be used for an accurate assessment of the vulnerability of typical bridges in the future.

The long-lasting co-evolution of ticks with pathogens results in mutual adaptation. Blood-feeding is one of the critical physiological behaviors that have been associated with the tick microbiome; however, most knowledge was gained through the study of laboratory-reared ticks. Here we detached Ixodes persulcatus ticks at different stages of blood-feeding from human patients and performed high-throughput transcriptomic analysis on them to identify their virome and genes differentially expressed between flat and fully fed ticks. We also traced bloodmeal sources of those ticks and identified bats and three other potential mammalian hosts, highlighting the public health significance. We found Jingmen tick virus and 13 putative new viruses belonging to 11 viral families, three of which even exhibited high genetic divergence from viruses previously reported in the same tick species from the same geographic region. Furthermore, differential expression analysis suggested a downregulation of antioxidant genes in the fully fed I. persulcatus ticks, which might be related to bloodmeal-related redox homeostasis. Our work highlights the significance of active surveillance of tick viromes and suggests a role of reactive oxygen species (ROS) in modulating changes in the microbiome during blood-feeding.

Background: A tick-borne segmented RNA virus called Jingmen tick virus (JMTV) was recently identified, variants of which were detected in a non-human primate host and fatal patients with Crimean-Congo haemorrhagic fever. We investigated its infectivity and pathogenicity for humans. Methods: We obtained skin-biopsy, blood and serum samples from patients with tick bites, and used high-throughput sequencing, in situ hybridisation, and serologic testing to diagnose and ascertain the cases of JMTV infection. Findings: A JMTV strain was isolated from the tick Amblyomma javanense into an embryo-derived tick cell line. We obtained sustained passage of JMTV, and revealed that it was able to accumulate in salivary glands of experimentally infected ticks. Four JMTV-infected patients were identified by high-throughput sequencing of skin biopsies and blood samples. The virus replication in skin tissue was visualised by in situ hybridisation. The four patients all had an itchy or painful eschar at the site of tick bite, with or without lymphadenopathy. Immunohistochemical examination revealed remarkable local inflammation manifested as infiltration by neutrophils. Eight patients were identified by serological testing and showed more severe clinical manifestations. Two Ixodes persulcatus ticks detached from patients were positive for JMTV. All JMTV strains identified in this study formed a well-supported sub-lineage, distinct from those previously reported in China.InterpretationThe public significance of JMTV should be highly concerning due to its potential pathogenicity for humans and efficient transmission by potential ticks. Fund: China Natural Science Foundation, State Key Research Development Programme, and United Kingdom Biotechnology and Biological Sciences Research Council. Keywords: Jingmen tick virus, Pathogenicity, Human infection, Ticks

The Yangtze–Huai River Basin (YHRB) always suffers from anomalously heavy rainfall during the warm season, and has been well explored as a whole area during the past several decades. In this study, the YHRB is divided into two core regions — the northern YHRB (nYHRB) and southern YHRB (sYHRB) — based on 29-year (1979–2007) June–July–August (JJA) temporally averaged daily rainfall rates and the standard deviation of rainfall. A spectral analysis of JJA daily rainfall data over these 29 years reveals that a 3–7-day synoptic-timescale high-frequency mode is absolutely dominant over the nYHRB, with 10–20-day and 15–40-day modes playing a secondary role. By contrast, 3–7-day and 10–20-day modes are both significant over the sYHRB, with 7–14-day, 15–40-day, and 20–60-day modes playing secondary roles. Based on a comparison between bandpass-filtered rainfall anomalies and original rainfall series, a total of 42, 1, 5, and 3 heavy rainfall events (daily rainfall amounts in the top 5% of rainy days) are detected over the nYHRB, corresponding to 3–7-day, 7–14-day, 10–20-day, and 15–40-day variation disturbances. Meanwhile, a total of 28, 8, 12, and 6 heavy rainfall events are detected over the sYHRB, corresponding to 3–7-day, 7–14-day, 10–20-day, and 20–60-day variation disturbances. The results have important implications for understanding the duration of summer heavy rainfall events over both regions.

We conducted an investigation of Borrelia miyamotoi infections in humans and ticks in northeastern China. Of 984 patients reporting recent tick bites, 14 (1.4%) were found to be infected with B. miyamotoi by PCR and genomic sequencing. The 14 patients had nonspecific febrile manifestations, including fever, headache, anorexia, asthenia, and arthralgia. Rash, eschar, and regional lymphadenopathy were each observed in 1 patient. Four (28.6%) patients were hospitalized because of severe disease. B. miyamotoi was detected in 3.0% (19/627) of Ixodes persulcatus, 1 (2.8%) of 36 Haemaphysalis concinna, and none of 29 Dermacentor silvarum ticks. Phylogenetic analyses based on sequences of a nearly entire 16s rRNA gene, a partial flagellin gene, and the glycerophosphodiester phosphodiesterase gene revealed that B. miyamotoi identified in patients and ticks were clustered in the group of the Siberian type. These findings indicate that B. miyamotoi is endemic in northeastern China and its public health significance deserves further investigation.

The tick-borne bacterium Anaplasma ovis is a widely distributed pathogen affecting sheep, goats and wild ruminants. Here, the HL-60 human promyelocytic leukemia cell line was used to isolate A. ovis from PCR-positive sheep and goats in Heilongjiang Province, China. Two weeks after inoculation, morulae were observed in cytoplasmic vacuoles in four different HL-60 cultures. Confocal microscopy using a Cy3-labeled A. ovis-specific probe confirmed that the HL-60 cells were infected with A. ovis. Cells from the 6th HL-60 subculture displayed positive fluorescence when incubated with A. ovis antiserum in the indirect fluorescent antibody assay. PCR amplification and sequencing of 16S rRNA, groEL, gltA, msp2 and msp4 Anaplasma genes revealed that the four A. ovis culture isolates were identical. Phylogenetic analysis showed that the sequences clustered with other A. ovis strains but could clearly be distinguished from other Anaplasma species. When the 18th subculture of infected HL-60 cells was examined by electron microscopy, lysosomes were often observed near the vacuoles. After the 24th subculture, Giemsa staining and PCR indicated that the HL-60 cells were negative for A. ovis. Although A. ovis can infect HL-60 cells for only four months, the ability of the organism to infect and multiply in HL-60 cells provides a tool to study intra-erythrocytic Anaplasma and host cell interactions.Emerging Microbes & Infections (2017) 6, e83; doi:10.1038/emi.2017.70; published online 20 September 2017

BACKGROUND:Babesia, usually found in wild and domestic mammals worldwide, have recently been responsible for emerging malaria-like zoonosis in infected patients. Human B. microti infection has been identified in China, primarily in the Southwest along the Myanmar border but little direct surveillance of B. microti infection in rodents has been carried out here (Yunnan province). In this region, a diverse topographic range combined with tropical moisture sustains a high biodiversity of small mammals, which might play important role on Babesia transmission. METHODS:Small mammals were captured in 141 sample locations from 18 counties located Yunnan Province, and screened for B. microti-like parasites infection by a nested PCR to target 18S rRNA gene of Babesia, plus directly sequencing for positive samples. Univariate and multivariate forward stepwise logistic regression analysis was used to access the association between infections and some related risk factors. RESULTS:Infection with Babesia microti was confirmed in 2.4% (53/ 2204) of small mammals. Significant differences in prevalence rates of B. microti were observed based on variations in forest, agricultural, and residential landscapes. Furthermore, adult small mammals had higher prevalence rates than younger, pubertal mammals. The near full-length 18S rRNA gene revealed that there were two types of B. microti, Kobe and Otsu, which demonstrate the genetic diversity and regional distribution. CONCLUSIONS:There exists a wide distribution and genetic diversity of endemic B. microti in Southwestern China, warranting further investigations and monitoring of clinical disease in individuals presenting with Babesia like symptoms in these areas.

The complete mitochondrial genome of Haemaphysalis concinna is reported for the first time in this study. Its entire mitogenome is 14,675 bp in length, contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and two non-coding regions. Among the 13 protein-coding genes, apart from the nad1, nad4, nad4L, and nad5 gene encoded on the L-strand, the remaining protein-coding genes were encoded on the H-strand. The phylogenetic analysis by Bayesian inference method shows that Amblyomma sphenodonti and H. parva formed one clade, while H. concinna and other species of genus Haemaphysalis formed the other clade, indicating that H. concinna belong to the genus Haemaphysalis.