Isolation of SARS-CoV-2-related coronavirus from Malayan pangolins.

The current outbreak of coronavirus disease-2019 (COVID-19) poses unprecedented challenges to global health1. The new coronavirus responsible for this outbreak-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-shares high sequence identity to SARS-CoV and a bat coronavirus, RaTG132. Although bats may be the reservoir host for a variety of coronaviruses3,4, it remains unknown whether SARS-CoV-2 has additional host species. Here we show that a coronavirus, which we name pangolin-CoV, isolated from a Malayan pangolin has 100%, 98.6%, 97.8% and 90.7% amino acid identity with SARS-CoV-2 in the E, M, N and S proteins, respectively. In particular, the receptor-binding domain of the S protein of pangolin-CoV is almost identical to that of SARS-CoV-2, with one difference in a noncritical amino acid. Our comparative genomic analysis suggests that SARS-CoV-2 may have originated in the recombination of a virus similar to pangolin-CoV with one similar to RaTG13. Pangolin-CoV was detected in 17 out of the 25 Malayan pangolins that we analysed. Infected pangolins showed clinical signs and histological changes, and circulating antibodies against pangolin-CoV reacted with the S protein of SARS-CoV-2. The isolation of a coronavirus from pangolins that is closely related to SARS-CoV-2 suggests that these animals have the potential to act as an intermediate host of SARS-CoV-2. This newly identified coronavirus from pangolins-the most-trafficked mammal in the illegal wildlife trade-could represent a future threat to public health if wildlife trade is not effectively controlled.

Therapeutic potential of Astragalus membranaceus-Pueraria lobata decoction for the treatment of chemotherapy bowel injury.

Intestinal mucositis (IM) is one of the most serious side effects of the chemotherapeutic agent irinotecan (CPT-11). Astragalus membranaceus-Pueraria lobata decoction is from the ancient medical book Zhengzhihuibu, has been reported to be used for the treatment of diabetes and hypertension. However, the beneficial effect and mechanism of AP on chemotherapy intestinal mucositis (CIM) remain largely unknown. This study aimed to investigate the efficacy and mechanism of Astragalus membranaceus-Pueraria lobata decoction (AP) in treating CIM. The beneficial effect and mechanism of AP on chemotherapy intestinal mucositis (CIM) were detected using Drosophila model, and combination with RT qPCR, transcriptomics. AP supplementation could significantly alleviate the CPT-11-induced body injury in Drosophila, such as increasing the survival rate, recovering the impaired digestion, improving the movement, and repairing the reproduction and developmental processes. Administration of AP remarkably alleviated the IM caused by CPT-11, including inhibiting the excretion, repairing the intestinal atrophy, improving the acid-base homeostasis imbalance, and inhibiting the disruption of intestinal structure. Mechanistic studies revealed that the protective role of AP against CPT-11 induced intestinal injury was regulated mainly by inhibiting immune-related Toll and Imd pathways, and enhancing the antioxidant capacity. Taken together, these results suggest that AP may be a novel agent to relieve CIM.

Assessment of nanotoxicity in a human placenta-on-a-chip from trophoblast stem cells.

Maternal exposure to nanoparticles during gestation poses potential risks to fetal development. The placenta, serving as a vital interface for maternal-fetal interaction, plays a pivotal role in shielding the fetus from direct nanoparticle exposure. However, the impact of nanoparticles on placental function is still poorly understood, primarily due to the absence of proper human placental models. In this study, we established a placenta-on-a-chip model capable of recapitulating nanoparticle exposure to assess potential nanotoxicity. The model was assembled by coculturing human trophoblast stem cells (hTSCs) and endothelial cells within a dynamic microsystem. hTSCs exhibited progressive differentiation into syncytiotrophoblasts under continuous fluid flow, forming a bilayered trophoblastic epithelium that mimicking both structural and functional aspects of human placental villi. Copper oxide nanoparticles (CuO NPs) were introduced into the trophoblastic side to simulate maternal blood exposure. Our findings revealed that CuO NPs hindered hTSCs differentiation, leading to diminished hormone secretion and impaired glucose transport. Subsequent analysis indicated that CuO NPs disrupted the autophagic flux in trophoblasts and induced apoptosis. Furthermore, the placenta-on-a-chip model exhibited inflammatory responses to CuO NP exposure, including maternal macrophage activation, inflammatory cytokine secretion, and endothelial barrier disruption. Dysfunction of the placental barrier and the ensuing inflammatory cascades may contribute to aberrant fetal development. Overall, our placenta-on-a-chip model offers a promising platform for assessing nanoparticle exposure-related risks and conducting toxicology studies.

Sympatric Recombination in Zoonotic Cryptosporidium Leads to Emergence of Populations with Modified Host Preference.

Genetic recombination plays a critical role in the emergence of pathogens with phenotypes such as drug resistance, virulence, and host adaptation. Here, we tested the hypothesis that recombination between sympatric ancestral populations leads to the emergence of divergent variants of the zoonotic parasite Cryptosporidium parvum with modified host ranges. Comparative genomic analyses of 101 isolates have identified seven subpopulations isolated by distance. They appear to be descendants of two ancestral populations, IIa in northwestern Europe and IId from southwestern Asia. Sympatric recombination in areas with both ancestral subtypes and subsequent selective sweeps have led to the emergence of new subpopulations with mosaic genomes and modified host preference. Subtelomeric genes could be involved in the adaptive selection of subpopulations, while copy number variations of genes encoding invasion-associated proteins are potentially associated with modified host ranges. These observations reveal ancestral origins of zoonotic C. parvum and suggest that pathogen import through modern animal farming might promote the emergence of divergent subpopulations of C. parvum with modified host preference.

Molecular Epidemiology of Human Cryptosporidiosis in Low- and Middle-Income Countries.

Cryptosporidiosis is one of the most important causes of moderate to severe diarrhea and diarrhea-related mortality in children under 2 years of age in low- and middle-income countries. In recent decades, genotyping and subtyping tools have been used in epidemiological studies of human cryptosporidiosis. Results of these studies suggest that higher genetic diversity of Cryptosporidium spp. is present in humans in these countries at both species and subtype levels and that anthroponotic transmission plays a major role in human cryptosporidiosis. Cryptosporidium hominis is the most common Cryptosporidium species in humans in almost all the low- and middle-income countries examined, with five subtype families (namely, Ia, Ib, Id, Ie, and If) being commonly found in most regions. In addition, most Cryptosporidium parvum infections in these areas are caused by the anthroponotic IIc subtype family rather than the zoonotic IIa subtype family. There is geographic segregation in Cryptosporidium hominis subtypes, as revealed by multilocus subtyping. Concurrent and sequential infections with different Cryptosporidium species and subtypes are common, as immunity against reinfection and cross protection against different Cryptosporidium species are partial. Differences in clinical presentations have been observed among Cryptosporidium species and C. hominis subtypes. These observations suggest that WASH (water, sanitation, and hygiene)-based interventions should be implemented to prevent and control human cryptosporidiosis in low- and middle-income countries.

Expression of Shh, Gli1, and Cyr61 in Gastric Cancer Predicts Overall Survival of Patients: A Retrospective Study.

OBJECTIVE: This study aimed to evaluate the expression levels of Shh, Gli1, and Cyr61 proteins in gastric cancer tissues and analyze the relationship between these three proteins and the clinicopathological factors and prognosis of patients. METHODS: This was a retrospective study. Four hundred gastric cancer tissue specimens from patients who underwent radical gastrectomy in Zhangye People's Hospital affiliated to Hexi University between February 2013 and February 2021 underwent immunohistochemical analysis. RESULTS: The positive expression rates of Shh, Gli1, and Cyr61 in gastric cancer tissues were 55.5%, 56.5%, and 64.5%, respectively. The expressions of Shh, Gli1, and Cyr61 in gastric cancer tissues were significantly correlated with tumor size, depth of invasion, and degree of differentiation (P < .05). The expression of Shh protein was positively correlated with the expression of Gli1 protein (P < .01), and the expression of Gli1 protein was positively correlated with the expression of Cyr61 protein (P < .01). Univariate and multivariate analyses showed that the expression of Shh, Gli1, and Cyr61 could predict the prognosis of patients (P < .05). Receiver operating characteristic curve analysis combined with TNM staging could better predict the three-year overall survival of patients (P < .05). CONCLUSION: Shh, Gli1, and Cyr61 proteins are significantly expressed in gastric cancer tissues and are risk factors for the prognosis of patients with gastric cancer.

Autologous platelet-rich plasma intrauterine perfusion to improve pregnancy outcomes after implantation failure: A systematic review and meta-analysis.

AIMS: Previous studies have reported inconsistent findings on the efficacy of platelet-rich plasma (PRP) therapy in women with implantation failure. The objective of this review was to evaluate whether PRP administration could improve pregnancy outcomes in women with implantation failure undergoing in vitro fertilization. METHODS: Electronic databases were searched for studies that explored the effects of PRP for patients with implantation failure. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. Based on the available data, we performed subgroup analyses and sensitivity analyses. RESULTS: Eight studies were included. PRP treatment improved pregnancy outcomes for all women compared with no treatment or placebo (clinical pregnancy rate: OR 2.24, 95% CI 1.41-3.54; live birth rate: OR 5.76, 95% CI 1.55-21.44; miscarriage rate: OR 0.18, 95% CI 0.05-0.63), especially in randomized controlled trials. No significant differences were detected in multiple pregnancy rates (OR 2.54, 95% CI 0.67-9.67). Furthermore, subgroup analysis based on the number of previous implantation failures showed that PRP treatment improved pregnancy outcomes in women with recurrent implantation failure (clinical pregnancy rate: OR 2.55, 95% CI 1.49-4.38; live birth rate: OR 5.07, 95% CI 1.15-22.34; miscarriage rate: OR 0.20, 95% CI 0.05-0.78). CONCLUSION: PRP administration could improve pregnancy outcomes in women with recurrent implantation failure. Due to the limited evidence available, the efficacy of PRP in women with recurrent implantation failure needs to be further verified in high-quality studies with larger sample sizes.

Characterization of Dense Granule Metalloproteinase INS-16 in Cryptosporidium parvum.

The protozoan pathogen Cryptosporidium parvum infects intestinal epithelial cells and causes diarrhea in humans and young animals. Among the more than 20 genes encoding insulinase-like metalloproteinases (INS), two are paralogs with high sequence identity. In this study, one of them, INS-16 encoded by the cgd3_4270 gene, was expressed and characterized in a comparative study of its sibling, INS-15 encoded by the cgd3_4260 gene. A full-length INS-16 protein and its active domain I were expressed in Escherichia coli, and antibodies against the domain I and an INS-16-specific peptide were produced in rabbits. In the analysis of the crude extract of oocysts, a ~60 kDa fragment of INS-16 rather than the full protein was recognized by polyclonal antibodies against the specific peptide, indicating that INS-16 undergoes proteolytic cleavage before maturation. The expression of the ins-16 gene peaked at the invasion phase of in vitro C. parvum culture, with the documented expression of the protein in both sporozoites and merozoites. Localization studies with antibodies showed significant differences in the distribution of the native INS-15 and INS-16 proteins in sporozoites and merozoites. INS-16 was identified as a dense granule protein in sporozoites and macrogamonts but was mostly expressed at the apical end of merozoites. We screened 48 candidate INS-16 inhibitors from the molecular docking of INS-16. Among them, two inhibited the growth of C. parvum in vitro (EC50 = 1.058 µM and 2.089 µM). The results of this study suggest that INS-16 may have important roles in the development of C. parvum and could be a valid target for the development of effective treatments.

Asymmetric and time-varying linkages between carbon emissions, globalization, natural resources and financial development in China.

In the real world, economic covariates follow asymmetric and time-varying patterns. Therefore, it is imperative to integrate these effects while estimating environmental and economic relationships. Although prevailing literature reveals various emissions-deriving and eliminating factors, however, there is a dearth of empirical evidence that estimates the asymmetric and time-varying effect of globalization, natural resources, and financial development from a multidimensional perspective in China. In doing so, we employ the nonlinear autoregressive distributed lag (NARDL) and cross-wavelet modeling framework to explore the long- and short-run nonlinear and time-variant association between globalization, natural resources, financial development, and carbon emissions from 1980 to 2017. The NARDL method has the benefit of discriminating the long-term and short-term asymmetric carbon emission responses due to a positive and negative shock in our primary variables of interest. Mainly, the findings of NARDL estimations confirm that positive shocks in globalization and financial developments have a significant positive impact on carbon emissions, whereas negative shock in natural resources has a significant positive impact on carbon emissions. Similarly, the outcomes of continuous wavelet transformation and wavelet transformation coherence confirm the causal linkages between covariates; however, this effect varies across different time and frequency domains. These results imply that environmental researchers should consider asymmetric transmission channels and time-frequency associations among variables to devise long-term sustainable policies.

Development of a Subtyping Tool for Zoonotic Pathogen Cryptosporidium canis.

Cryptosporidium canis is an important cause of cryptosporidiosis in canines and humans. Studies of the transmission characteristics of C. canis are currently hampered by the lack of suitable subtyping tools. In this study, we conducted a genomic survey of the pathogen and developed a subtyping tool targeting the partial 60-kDa glycoprotein gene (gp60). Seventy-six isolates previously identified as C. canis were analyzed using the new subtyping tool. Amplicons of the expected size were obtained from 49 isolates, and phylogenetic analysis identified 10 subtypes clustered into five distinct groups (XXa to XXe). The largest group, XXa, contained 43 isolates from four subtypes that differed slightly from each other at the nucleotide level, while groups XXb to XXe contain one to three isolates each. The similar distributions of subtypes in humans and canines suggest that zoonotic transmission might play an important role in the epidemiology of C. canis In addition, suspected zoonotic transmission of C. canis between dogs and humans in a household was confirmed using the subtyping tool. The subtyping tool and data generated in this study might improve our understanding of the transmission of this zoonotic pathogen.

Diversity and emendation of the genus Planktoniella (Bacillariophyceae), with descriptions of P. tubulata sp. nov. and P. trifurcata sp. nov.

For the first time, LSU and SSU sequences were obtained from four species of the marine planktonic diatom Planktoniella. Samples were collected and cultured from Chinese coastal waters, and morphological observations were made using light and scanning electron microscopy. Combined morphological and DNA sequence data revealed two new species herein described as Planktoniella tubulata and Planktoniella trifurcata. Planktoniella tubulata is characterized by the prominent external tubes of two rimoportulae and the long external tubes of the marginal fultoportulae. The latter are organized regularly in a ring, externally on the mantle, and protrude parallel to the valvar plane. Planktoniella trifurcata is unique in having mucilaginous extensions that usually expand into three lobes, and external extensions of the marginal fultoportulae being either tubes or spines. In the phylogenetic analyses inferred from SSU and LSU sequences, the four analyzed Planktoniella taxa formed a well-supported monophyletic group. A suite of three characters, mucilage membranes or wings, two marginal rimoportulae, and mantle ribs are diagnostic for Planktoniella. Based on these detailed ultrastructural observations, the description of Planktoniella is emended.

Evaluation of hepatic drug-metabolism for glioblastoma using liver-brain chip.

Glioma is one of the most aggressive and highly fatal diseases with an extremely poor prognosis. Considering the poor clinical response to therapy in glioma, it is urgent to establish an in vitro model to facilitate the screening and assessment of anti-brain-tumor drugs. The blood-brain barrier (BBB), as well as liver metabolism plays an important role in determining the pharmacological activity of many anti-brain-tumor drugs. In this work, we designed a multi-interface liver-brain chip integrating co-culture system to assess hepatic metabolism dependent cytotoxicity of anti-brain-tumor drug in vitro. This microdevice composed of three microchannels which were separated by porous membrane and collagen. HepG2 and U87 cells were cultured in separated channels as mimics of liver and glioblastoma. Brain microvascular endothelial cells (BMECS) and cerebral astrocytes were co-cultured on collagen to mimic the brain microvascular endothelial barrier. Three common anti-tumor drugs, paclitaxel (PTX), capecitabine (CAP) and temozolomide (TMZ), were evaluated on this chip. In integrated liver-brain chip, liver enhanced the cytotoxicity of CAP on U87 cells by 30%, but having no significant effect on TMZ. The BBB decreased the cytotoxicity of PTX by 20%, while no significant effects were observed on TMZ and CAP, indicating the importance of liver metabolism and blood-brain barrier on the evaluation of anti-brain-tumor drugs. This work provides a biomimetic liver-brain model to mimic the physiological and pharmacological processes in vitro and presents a simple platform for long-term cell co-culture, drug delivery and metabolism, and real-time analysis of drug effects on brain cancer.

Small ruminants and zoonotic cryptosporidiosis.

Sheep and goats are commonly infected with three Cryptosporidium species, including Cryptosporidium parvum, Cryptosporidium ubiquitum, and Cryptosporidium xiaoi, which differ from each in prevalence, geographic distribution, and public health importance. While C. parvum appears to be a dominant species in small ruminants in European countries, its occurrence in most African, Asian, and American countries appear to be limited. As a result, zoonotic infections due to contact with lambs and goat kids are common in European countries, leading to frequent reports of outbreaks of cryptosporidiosis on petting farms. In contrast, C. xiaoi is the dominant species elsewhere, and mostly does not infect humans. While C. ubiquitum is another zoonotic species, it occurs in sheep and goats at much lower frequency. Host adaptation appears to be present in both C. parvum and C. ubiquitum, consisting of several subtype families with different host preference. The host-adapted nature of C. parvum and C. ubiquitum has allowed the use of subtyping tools in tracking infection sources. This has led to the identification of geographic differences in the importance of small ruminants in epidemiology of human cryptosporidiosis. These tools have also been used effectively in linking zoonotic transmission of C. parvum between outbreak cases and the suspected animals. Further studies should be directly elucidating the reasons for differences in the distribution and public health importance of major Cryptosporidium species in sheep and goats.

Appraising the Value of Serum and Serum-Derived Exosomal LncRNA-EXOC7 as a Promising Biomarker in Cervical Cancer.

BACKGROUND: The abnormally expressed long non-coding RNAs (lncRNAs) are closely related to the onset and progression of various malignant tumors. In this study, we aimed to explore the value of serum and serum-derived exosomal lncRNA-EXOC7 (long non-coding RNA exocyst complex component 7) in the diagnosis and monitoring of cervical cancer (CC). METHODS: The expression of lncRNA-EXOC7 in serum and serum-derived exosomes in CC patients was detected by quantitative real-time PCR (qRT-PCR), and the correlations between lncRNA-EXOC7 expression and clinicopathological characteristics were analyzed by Spearman's and Chi-square tests. RESULTS: Serum vesicles were successfully isolated and identified. The expression of lncRNA-EXOC7 in serum and serum-derived exosomes in CC patients was markedly elevated compared with that in healthy controls. The AUCs of serum and exosomal lncRNA-EXOC7 in distinguishing CC patients from healthy controls were 0.9388 and 0.8982, respectively. The expression of lncRNA-EXOC7 in serum and exosomes was correlated with the FIGO stage of CC (p = 0.041 or 0.010), and positively correlated with levels of CYFRA211, TPS, and SCC (all with p < 0.05). Serum and exosomal lncRNA-EXOC7 was related to the treatment and recurrence of CC; that means, it was significantly repressed after treatment and up-regulated at the time of recurrence. CONCLUSIONS: Serum and exosomal lncRNA-EXOC7 can be used as an important biomarker for the diagnosis, the evaluation of curative effect and the detection of recurrence of CC.

Zoonotic potential of Enterocytozoon bieneusi and Giardia duodenalis in horses and donkeys in northern China.

Limited data are available on infection rates and genetic identity of Enterocytozoon bieneusi and Giardia duodenalis in horses and donkeys. In this study, 865 fecal specimens were collected from donkeys (n = 540) and horses (n = 325) in three provinces and autonomous regions in northern China during 2015-2019. Enterocytozoon bieneusi was detected and genotyped by PCR and sequence analyses of the ribosomal internal transcribed spacer (ITS) and G. duodenalis was detected and genotyped by PCR and sequence analyses of the ß-giardin, glutamate dehydrogenase, and triosephosphate isomerase genes. The overall infection rates of E. bieneusi and G. duodenalis were 21.9% (118/540) and 11.5% (62/540) in donkeys, and 7.4% (24/325) and 2.8% (9/325) in horses, respectively. These differences in infection rates of E. bieneusi and G. duodenalis between donkeys and horses were significant (χ2 = 30.9, df = 1, P < 0.0001; χ2 = 20.4, df = 1, P < 0.0001, respectively). By age, the 28.9% infection rate of E. bieneusi in donkeys under 6 months was significantly higher than that in animals over 6 months (6.0%; χ2 = 35.2, df = 1, P < 0.0001). In contrast, donkeys of 6-12 months had higher infection rate (35.9%) of G. duodenalis than donkeys under 6 months (9.9%; χ2 = 22.1, df = 1, P < 0.0001) and over 12 months (8.7%; χ2 = 17.3, df = 1, P < 0.0001). In horses, animals of > 12 months had significantly higher infection rate (31.1%) of E. bieneusi than horses under 6 months (3.4%; χ2 = 29.4, df = 1, P < 0.0001) and 6-12 months (3.8%; χ2 = 26.1, df = 1, P < 0.0001). Twenty genotypes of E. bieneusi were detected, including six known ones and 14 new genotypes. Among them, nine genotypes in 45% E. bieneusi-positive specimens belonged to the zoonotic group 1. Similarly, three G. duodenalis assemblages were detected, including A (in 2 horses and 30 donkeys), B (in 6 horses and 29 donkeys), and E (in 1 horse); three donkeys had coinfections of assemblages A and B. The assemblage A isolates identified all belong to the sub-assemblage AI. These results indicate that unlike in other farm animals, there is a common occurrence of zoonotic E. bieneusi and G. duodenalis genotypes in horses and donkeys.

Contribution of hospitals to the occurrence of enteric protists in urban wastewater.

We assessed the potential contribution of hospitals to contaminations of wastewater by enteric protists, including Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in raw wastewater. Wastewater samples were collected from storage tanks in two hospitals and one associated wastewater treatment plant in Shanghai, China, from March to November 2009. Enteric pathogens were detected and identified using PCR and DNA sequencing techniques. Among a total of 164 samples analyzed, 31 (18.9%), 45 (27.4%), and 122 (74.4%) were positive for Cryptosporidium spp., G. duodenalis, and E. bieneusi, respectively. Altogether, three Cryptosporidium species, four G. duodenalis assemblages, and 12 E. bieneusi genotypes were detected. Cryptosporidium hominis, G. duodenalis sub-assemblage AII, and E. bieneusi genotype D were the dominant ones in wastewater from both hospitals and the wastewater treatment plant. A similar distribution in genotypes of enteric pathogens was seen between samples from hospitals and the wastewater treatment plant, suggesting that humans are one of the major sources for these pathogens and hospitals are important contributors of enteric parasites in urban wastewater. Data from this study might be useful in the formulation of preventive measures against environmental contamination of waterborne pathogens.

Comparative analysis reveals conservation in genome organization among intestinal Cryptosporidium species and sequence divergence in potential secreted pathogenesis determinants among major human-infecting species.

BACKGROUND: Cryptosporidiosis is a major cause of gastrointestinal diseases in humans and other vertebrates. Previous analyses of invasion-related proteins revealed that Cryptosporidium parvum, Cryptosporidium hominis, and Cryptosporidium ubiquitum mainly differed in copy numbers of secreted MEDLE proteins and insulinase-like proteases and sequences of mucin-type glycoproteins. Recently, Cryptosporidium chipmunk genotype I was identified as a novel zoonotic pathogen in humans. In this study, we sequenced its genome and conducted a comparative genomic analysis. RESULTS: The genome of Cryptosporidium chipmunk genotype I has gene content and organization similar to C. parvum and other intestinal Cryptosporidium species sequenced to date. A total of 3783 putative protein-encoding genes were identified in the genome, 3525 of which are shared by Cryptosporidium chipmunk genotype I and three major human-pathogenic Cryptosporidium species, C. parvum, C. hominis, and Cryptosporidium meleagridis. The metabolic pathways are almost identical among these four Cryptosporidium species. Compared with C. parvum, a major reduction in gene content in Cryptosporidium chipmunk genotype I is in the number of telomeric genes encoding MEDLE proteins (two instead of six) and insulinase-like proteases (one instead of two). Highly polymorphic genes between the two species are mostly subtelomeric ones encoding secretory proteins, most of which have higher dN/dS ratios and half are members of multiple gene families. In particular, two subtelomeric ABC transporters are under strong positive selection. CONCLUSIONS: Cryptosporidium chipmunk genotype I possesses genome organization, gene content, metabolic pathways and invasion-related proteins similar to the common human-pathogenic Cryptosporidium species, reaffirming its human-pathogenic nature. The loss of some subtelomeric genes encoding insulinase-like proteases and secreted MEDLE proteins and high sequence divergence in secreted pathogenesis determinants could contribute to the biological differences among human-pathogenic Cryptosporidium species.

Infection patterns, clinical significance, and genetic characteristics of Enterocytozoon bieneusi and Giardia duodenalis in dairy cattle in Jiangsu, China.

The infection patterns and clinical significance of Enterocytozoon bieneusi and Giardia duodenalis in dairy cattle remain poorly investigated despite their common occurrence. Data on the genetic diversity are also needed to understand the transmission and human-infective potential of the two pathogens. In this study, fecal specimens from 1366 dairy cattle on a large farm were examined for the presence and genotype distribution of E. bieneusi and G. duodenalis by PCR and DNA sequencing. The overall infection rates of E. bieneusi and G. duodenalis were 13.0% and 20.6%, respectively. Pre-weaned calves had significantly higher infection rates of both pathogens than post-weaned and adult cattle (P < 0.001), with peak occurrence of the pathogens in animals of 7-12 weeks. In both pre- and post-weaned calves, animals with diarrhea were 2.1-3.0 times more likely to be infected with either pathogen than those without diarrhea (P < 0.01). The E. bieneusi identified belonged to five genotypes, including J (n = 138), I (n = 21), BEB4 (n = 10), Type IV (n = 1), and a novel genotype CHC17 (n = 1). Genotype J was the dominant one in all age groups, whereas genotype I was only identified in calves of 6-11 weeks. Genotyping of G. duodenalis at three genetic loci identified assemblage E (n = 278), assemblage A (n = 2), and concurrence of the two (n = 1). Altogether, 13, 7 and 10 subtypes of assemblage E were detected at the bg, gdh, and tpi loci, respectively, forming 65 multilocus genotypes. The formation of two major clusters of MLGs in eBURST analysis indicated that intra-assemblage genetic recombination of two dominant MLGs could have led to the high genetic heterogeneity within assemblage E on a single farm. Results of this study provide much needed data on the pathogenicity of E. bieneusi and G. duodenalis in pre- and post-weaned calves. The clinical significance of the two pathogens in dairy cattle warrants further investigations.

A Biomimetic Human Gut-on-a-Chip for Modeling Drug Metabolism in Intestine.

Drug metabolism in the intestine is considered to substantially contribute to the overall first-pass metabolism, which has been neglected for a long time. It is highly desirable to develop a reliable model to evaluate drug metabolism in the intestine in vitro. In this work, we made the first attempt to develop a biomimetic human gut-on-a-chip for modeling drug metabolism in intestine. In this chip, constant flow, together with porous nitrocellulose membrane and collagen I, mimics an in vivo-like intestinal microenvironment. The Caco-2 cells grown in the chip formed a compact intestinal epithelial layer with continuous expression of the tight junction protein, ZO-1. Furthermore, higher gene expression of villin, sucrase-isomaltase, and alkaline phosphatase demonstrated that cells in the biomimetic human gut-on-a-chip device were more mature with near-physiological functions compared to the control on planar substrate. In particular, cellular metabolic activity was assessed on different substrates, indicating higher metabolic efficiency of ifosfamide and verapamil in the biomimetic human gut-on-a-chip model. Taken together, our results suggested that this biomimetic human gut-on-a-chip promoted the differentiation of intestinal cells with enhanced functionality by creating a biomimetic 3D microenvironment in vitro. It might offer a bioactive, low-cost, and flexible in vitro platform for studies on intestinal metabolism as well as preclinical drug development.