Health Services Utilization in China during the COVID-19 Pandemic: Results from a Large-Scale Online Survey

Timely access to essential health services is a concern as COVID-19 continues. This study aimed to investigate health services utilization during the first wave of the pandemic in China. A cross-sectional online survey was conducted using a self-administrated questionnaire in March 2020. Descriptive statistics and logistic regression were used for data analysis. A total of 4744 respondents were included, with 52.00% reporting affected services utilization. Clinical testing (68.14%) and drug purchase (49.61%) were the most affected types. Higher education level, being married, chronic disease, frequently visiting a provincial medical institution, spending more time on pandemic-related information, perception of high-risk of infection, perception of large health impact of the pandemic, and anxiety/depression were significant predictors for reporting affected services utilization. For the 431 chronic disease respondents, 62.18% reported interruption, especially for drug purchase (58.58%). Affected health services utilization was reported during the first wave of the pandemic in China, especially for those with higher education level, chronic diseases, and COVID-19 related concerns. Enhancing primary healthcare, use of telehealth, extended prescription, and public communication were countermeasures undertaken by China during the rapid rise period. As COVID-19 progresses, the changing disease characteristics, adapted health system, along with enhanced public awareness/knowledge should be considered for the evolution of health services utilization, and further investigation is needed

A high-throughput screen identifies inhibitors of lung cancer stem cells

Metastasis is the main cause of cancer morbidity and mortality. Cancer stem cells (CSCs) are a rare subpopulation of cancer cells that can drive metastasis. The identification of CSC inhibitors and CSC-related genes is an alluring strategy for suppressing metastasis. Here, we established a simple and repeatable high-throughput CSC inhibitor screening platform that combined tumor sphere formation assays and cell viability assays. Human lung cancer cells were cocultured with 1280 pharmacologically active compounds (FDA-approved). Fifty-four candidate compounds obtained from our screening system completely or partially inhibited tumor sphere formation. A total of 5 of these 54 compounds (prochlorperazine dimaleate, thioridazine hydrochloride, ciproxifan hydrochloride, Ro 25-6981 hydrochloride, and AMN 082) completely inhibited the self-renewal of CSCs without cytotoxicity in vitro via their targets and suppressed lung cancer metastasis in vivo, suggesting that our screening platform is selective and reliable. DRD2, HRH3, and GRIN2B exhibited potent genes promoting CSCs in vitro experiments and clinical datasets. Further validation of the top hit (DRD2) and previously published studies demonstrate that our screening platform is a useful tool for CSC inhibitor and CSC-related gene screening

A Fatal Pneumonia due to Coinfection of Pseudomonas putida and Staphylococcus pseudintermedius in a Laboratory Beagle Dog

Background: Pseudomonas putida (P. putida) is widely distributed in the environment, and sometimes caused nosocomial infections in human beings, but no case of infection has been reported in beagle dogs. Staphylococcus pseudintermedius (S. pseudintermedius) is a natural cutaneous bacterium in dogs and occasionally causes purulent infections of the skin and rarely causes pneumonia. Both bacteria are opportunistic pathogens. Dogs, even well-controlled laboratory beagle dogs, maybe infected by the bacterium in certain conditions like this report. In order to provide information and give suggestion to veterinarians involved in dogs study, a complete profile of the coinfection was drawn in this report.Case: It is presented a case of an 8-month-old beagle dog, weighing 6 kg that suffered from coinfection of P. putida and S. pseudintermedius during a treatment of chemotherapy. The animal was confirmed as normal by appearance, physical examination, and laboratory tests before arrival according to the applicable guidelines. After 14-day acclimation period, the animal was administrated with a tyrosinase inhibitor once daily via oral gavage. From Day 8, coughing, decreased activity, hyporeflexia, squinting, shortness of breath (abdominal breathing), and discharge around the nose as well as crackles in the lung and rapid heart rate were noted. Since the poor conditions progressed rapidly and have not been improved by treatment of ceftriaxone and dexamethasone. On Day 9, the animal was euthanized for humanitarian reasons due to rapid progress and poor condition. To define the pathogen, hilar lymph node and thoracic swab were collected for bacteria isolation and purification in special mediums, and at last characterized by Gram staining and 16s rRNA gene sequence analysis and positive PCR-restriction fragment length polymorphism. In clinical pathological examination, an increase in WBC, neutrophils, lymphocytes, monocytes, cholesterol, triglyceride, total protein, globulin, and lactate dehydrogenase, as well as a decrease in RBC, hemoglobin, hematocrit, platelets, sodium ion, chloride ion, and albumin were noted. At necropsy, dark red and enlarged lymph nodes were noted in the hilum of lung, multiple abscesses with yellow pus and multifocal hemorrhage were noted in the lung, and a large amount of frothy yellow fluid were noted in the trachea. In pathological examinations, severe neutrophilic inflammation, diffuse and moderate macrophage aggregation, mild hemorrhage, and moderate alveolar emphysema were noted in the lung, and severe sinusoidal stasis were noted in portal lymph nodes.Discussion: The current case presented a profile of the appearance, treatment, hematological examination, coagulation examination, clinical chemistry, macroscopic and histological changes in the lung. Multiple purulent abscesses, infiltration of neutrophils, macrophage, and hemorrhage, were correlated to the increase in WBC, neutrophils, lymphocytes, and monocytes, and the decrease in RBC, hemoglobin, hematocrit, and platelets. In the coagulation examination, an increase in Fbg concentration was noted. This change may be induced by the coagulase effect of the S. pseudintermedius, yet no effect on PT or APTT was noted, indicating the coagulation function has not been affected. In the clinical chemistry, the increase of creatine kinase and lactate dehydrogenase may indicate tissue cell damages. Significant increase of globulin may be caused by the inflammatory status. In conclusion, the findings in this case indicate that both Pseudomonas putida and Staphylococcus pseudintermedius can induce infections in laboratory beagle dogs under certain conditions, and might result in a fatal pneumonia which could progress very fast within several days

Genome-Wide Analysis of Sugar Transporters Identifies the gtsA Gene for Glucose Transportation in Pseudomonas stutzeri A1501

Pseudomonas stutzeri A1501 possesses an extraordinary number of transporters which confer this rhizosphere bacterium with the sophisticated ability to metabolize various carbon sources. However, sugars are not a preferred carbon source for P. stutzeri A1501. The P. stutzeri A1501 genome has been sequenced, allowing for the homology-based in silico identification of genes potentially encoding sugar-transport systems by using established microbial sugar transporters as a template sequence. Genomic analysis revealed that there were 10 sugar transporters in P. stutzeri A1501, most of which belong to the ATP-binding cassette (ABC) family (5/10); the others belong to the phosphotransferase system (PTS), major intrinsic protein (MIP) family, major facilitator superfamily (MFS) and the sodium solute superfamily (SSS). These systems might serve for the import of glucose, galactose, fructose and other types of sugar. Growth analysis showed that the only effective medium was glucose and its corresponding metabolic system was relatively complete. Notably, the loci of glucose metabolism regulatory systems HexR, GltR/GtrS, and GntR were adjacent to the transporters ABCMalEFGK, ABCGtsABCD, and ABCMtlEFGK, respectively. Only the ABCGtsABCD expression was significantly upregulated under both glucose-sufficient and -limited conditions. The predicted structure and mutant phenotype data of the key protein GtsA provided biochemical evidence that P. stutzeri A1501 predominantly utilized the ABCGtsABCD transporter for glucose uptake. We speculate that gene absence and gene diversity in P. stutzeri A1501 was caused by sugar-deficient environmental factors and hope that this report can provide guidance for further analysis of similar bacterial lifestyles

Molecular characterization of hemotropic mycoplasmas (Mycoplasma ovis and ‘Candidatus Mycoplasma haemovis’) in sheep and goats in China

Abstract Background Hemotropic mycoplasmas (hemoplasmas) are emerging zoonotic pathogens with a worldwide distribution that can cause mild to severe hemolytic anemia, icterus, ill-thrift, infertility, and poor weight gain. However, understanding of the molecular epidemiology of hemoplasmas (Mycoplasma ovis and ‘Candidatus Mycoplasma haemovis’) is limited in sheep and goats, and the hemoplasma strain/species/variant ‘Candidatus M. haemovis’ was poorly studied throughout the world and had never been detected in China until now. Thus, the aim of the present study was to determine the molecular prevalence of hemoplasmas, including M. ovis and ‘Candidatus M. haemovis’ in sheep and goats from seven provinces and one autonomous region of China. Methods A total of 1364 blood samples were collected from sheep and goats in seven provinces and one autonomous region of China. All blood samples were tested for hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) by nested PCR amplification based on 16S rRNA gene. Positive specimens underwent nucleotide sequencing and phylogenetic analysis. Results Overall, 610 specimens (44.7%, 610/1364) were shown to be hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) -positive by nested PCR amplification based on 16S rRNA gene. The prevalence in goats was 44.1% (379/860), and 45.8% (231/504) in sheep, while that in grazing small ruminants was 54.4% (396/728) and 33.6% (214/636) in house feeding small ruminants. Sequencing of the nearly complete 16S rRNA gene was successful for the 103 randomly selected positive specimens from different farms in different sampling sites of China. Among them, analysis of the 16S rRNA gene sequences identified M. ovis (n = 56) and ‘Candidatus M. haemovis’ (n = 47). Two (KU983740 and KU983746) of the four novel genotypes obtained in this study were closely related to M. ovis, while the other two genotypes (KU983748 and KU983749) had high identity with ‘Candidatus M. haemovis’. Remarkably, the genotype (KU983740) of M. ovis in sheep and goats in this study fell in a clade with two human hemoplasmas from USA (KF313922 and GU230144) and shared 99.8%–99.9% with them. Conclusions In this study, ‘Candidatus M. haemovis’ was first detected in Chinese sheep and goats and hemoplasmas (M. ovis and ‘Candidatus M. haemovis’) are highly prevalent, and widely distributed in China

The complete mitochondrial genome of Fannia scalaris (Diptera: Muscidae)

Fannia scalaris (Fabricius, 1794) is closely related to human life in ecological habits, which can lead to health concerns since they feed on various contamination sources. In this study, we first present the mitochondrial genome (mitogenome) of F. scalaris (GenBank No. MT017706). The length of this mitogenome was composed of 15,040 base pairs, including 13 protein-coding genes, two ribosomal RNA, 22 transfer RNA, and an AT-rich region. It consisted of A 39.3%, G 9.1%, C 13.0%, and T 38.6%. The arrangement of the genes was consistent with that of the ancestral metazoan. Furthermore, phylogenetic relationship indicated that F. scalaris was obviously separated from the muscid flies. This study provides useful genetic data in order to further understand the evolutionary relationship of the Muscidae

Cryptosporidium spp. in large-scale sheep farms in China: prevalence and genetic diversity

Abstract Cryptosporidium spp. are significant zoonotic intestinal parasites that induce diarrhea and even death across most vertebrates, including humans. Previous studies showed that sheep are important hosts for Cryptosporidium and that its distribution in sheep is influenced by geography, feeding patterns, age, and season. Environmental factors also influence the transmission of Cryptosporidium. Molecular studies of Cryptosporidium in sheep have been conducted in only a few regions of China, and studies into the effect of sheep-housing environments on Cryptosporidium transmission are even rarer. To detect the prevalence of Cryptosporidium in large-scale sheep-housing farms, a total of 1241 fecal samples were collected from sheep, 727 environmental samples were taken from sheep housing, and 30 water samples were collected in six regions of China. To ascertain the existence of the parasite and identify the species of Cryptosporidium spp., we conducted nested PCR amplification of DNA extracted from all samples using the small-subunit (SSU) rRNA gene as a target. For a more in-depth analysis of Cryptosporidium spp. subtypes, C. xiaoi-and C. ubiquitum-positive samples underwent separate nested PCR amplification targeting the 60 kDa glycoprotein (gp60) gene. The amplification of the Cryptosporidium spp. SSU rRNA gene locus from the whole genomic DNA of all samples yielded a positive rate of 1.2% (20/1241) in fecal samples, 0.1% (1/727) in environmental samples, and no positive samples were found in water samples. The prevalence of Cryptosporidium spp. infection in large-scale housed sheep was 1.7%, which was higher than that in free-ranging sheep (0.0%). The highest prevalence of infection was found in weaning lambs (6.8%). Among the different seasons, the peaks were found in the fall and winter. The most prevalent species were C. xiaoi and C. ubiquitum, with the former accounting for the majority of infections. The distribution of C. xiaoi subtypes was diverse, with XXIIIc (n = 1), XXIIId (n = 2), XXIIIe (n = 2), and XXIIIl (n = 4) identified. In contrast, only one subtype, XIIa (n = 9), was found in C. ubiquitum. In this study, C. xiaoi and C. ubiquitum were found to be the predominant species, and Cryptosporidium was found to be present in the environment. These findings provide an important foundation for the comprehensive prevention and management of Cryptosporidium in intensively reared sheep. Furthermore, by elucidating the prevalence of Cryptosporidium in sheep and its potential role in environmental transmission, this study deepens our understanding of the intricate interactions between animal health, environmental contamination, and public health dynamics

Fibroblast-Derived Exosomes Contribute to Chemoresistance through Priming Cancer Stem Cells in Colorectal Cancer.

Chemotherapy resistance observed in patients with colorectal cancer (CRC) may be related to the presence of cancer stem cells (CSCs), but the underlying mechanism(s) remain unclear. Carcinoma-associated fibroblasts (CAFs) are intimately involved in tumor recurrence, and targeting them increases chemo-sensitivity. We investigated whether fibroblasts might increase CSCs thus mediating chemotherapy resistance. CSCs were isolated from either patient-derived xenografts or CRC cell lines based on expression of CD133. First, CSCs were found to be inherently resistant to cell death induced by chemotherapy. In addition, fibroblast-derived conditioned medium (CM) promoted percentage, clonogenicity and tumor growth of CSCs (i.e., CD133+ and TOP-GFP+) upon treatment with 5-fluorouracil (5-Fu) or oxaliplatin (OXA). Further investigations exhibited that exosomes, isolated from CM, similarly took the above effects. Inhibition of exosome secretion decreased the percentage, clonogenicity and tumor growth of CSCs. Altogether, our findings suggest that, besides targeting CSCs, new therapeutic strategies blocking CAFs secretion even before chemotherapy shall be developed to gain better clinical benefits in advanced CRCs