Toxoplasma gondii infection is associated with schizophrenia from the perspectives of seroepidemiology and serum metabolomics in Hunan Province, China.

Toxoplasma gondii (T.gondii) can influence the host's neurotransmission, central immune responses, and brain structure, potentially impacting the onset and development of various psychiatric disorders such as schizophrenia. We employed Electrochemiluminescence Immunoassay (ECLIA) to measure anti-Toxoplasma antibodies in 451 schizophrenic patients and 478 individuals from the general population in Hunan, China. The incidence rate of T.gondii infection in schizophrenic patients (8.87 %) was higher than that in the general population (3.77 %). A significant difference was observed among females, but not in males. Age-stratified analysis revealed significant differences in the 21-40 and 41-60 age groups. The two populations had no significant difference in the antibody titer for T. gondii infection. Additionally, the profile of circulating metabolites in the serum of schizophrenic patients with or without T. gondii infection was examined using non-targeted metabolomics assay. A total of 68 metabolites were differentially expressed between Toxoplasma-positive and Toxoplasma-negative groups, potentially mediating the connection between T. gondii infection and schizophrenia. Our research suggests that schizophrenic patients are susceptible to T. gondii infection with distinct metabolic program.

Fatal, non-fatal burden of cancer in the elderly in China, 2005-2016: a nationwide registry-based study.

BACKGROUND: As populations age, cancer burden becomes increasingly conspicuous. This study quantified the cancer burden of the elderly (≥ 60 years) in China, based on the China Cancer Registry Annual Report to provide epidemiological evidence for cancer prevention and control. METHODS: Data on cancer cases and deaths among the elderly aged ≥ 60 years were collected from the China Cancer Registry Annual Report, 2008-2019. Potential years of life lost (PYLL) and disability-adjusted life years (DALY) were calculated to analyze fatalities and the non-fatal burden. The time trend was analyzed using the Joinpoint model. RESULTS: From 2005 to 2016, the PYLL rate of cancer in the elderly was stable between 45.34‰ and 47.62‰, but the DALY rate for cancer decreased at an average annual rate of 1.18% (95% CI: 0.84-1.52%). The non-fatal cancer burden in the rural elderly was higher than that of the urban elderly. Lung, gastric, liver, esophageal, and colorectal cancers were the main cancers causing the cancer burden in the elderly, and accounted for 74.3% of DALYs. The DALY rate of lung cancer in females in the 60-64 age group increased (annual percentage change [APC] = 1.14%, 95% CI: 0.10-1.82%). Female breast cancer was one of the top five cancers in the 60-64 age group, with DALY rates that also increased (APC = 2.17%, 95% CI: 1.35-3.01%). With increasing age, the burden of liver cancer decreased, while that of colorectal cancer rose. CONCLUSIONS: From 2005 to 2016, the cancer burden in the elderly in China decreased, mainly reflected in the non-fatal burden. Female breast and liver cancer were a more serious burden in the younger elderly, while colorectal cancer burden was mainly observed in the older elderly.

Bioinformatics of excretory/secretory proteins of Toxoplasma gondii strain ME49.

Toxoplasma gondii is an obligate intracellular parasite that is globally distributed and can infect almost all warm-blooded animals, including humans. While vaccines are not available for prophylaxis, there are limited therapeutic options which often do not result in eradication of parasites from patients. ME49 is a cystogenic strain of T. gondii with a potential as a vaccine candidate. Excretory and secretory (ES) proteins are thought to play crucial roles in host-parasite interactions. Hence, we predicted and functionally annotated the ES proteins in T. gondii ME49 using public databases. ES proteins were further examined for the characteristics and possible functions through gene ontology (GO) term enrichment and analyses of metabolic pathways, enzyme code distribution (EC distributing) and protein domains. The potential antigenicity of T. gondii ME49 ES proteins was evaluated for the first time using Abundance of Antigenic Regions (AAR) value to predict the antigenic potential by measuring sequence length and number of antigenic regions. The results showed that the ES proteins have different AAR values at RNA and microarray level. Our studies provide valuable information for future experimental studies, including the identification of novel ES proteins of therapeutic, diagnostic and immunoprophylactic interests.

Exposure time determines the protective effect of Trichinella spiralis on experimental colitis.

Several studies demonstrate the protective effect of Trichinella spiralis (T. spiralis) on autoimmune diseases, however the optimal exposure time remains unexplored. This study aimed to determine whether pre-exposure of mice to T. spiralis conferred greater protection than introduction of the parasite in the acute phase of experimental colitis. We compared the effect of T. spiralis on dextran sodium sulfate (DSS)-induced colitis using two exposure paradigms: introduction three weeks prior to, or immediately after the induction period. Inflammation scores, morphological changes and cytokine profiles in serum and colonic tissue were assessed. At a parasite dose of 300 cysts, post exposure had a more pronounced effect on cytokine profiles, improved gross appearance of colon tissue, and reduced inflammatory symptoms. In addition, we demonstrate that regardless of cyst number, pre-exposure to T. spiralis did not confer protective benefits when compared to parasite introduction in the acute phase of DSS-induced colitis. Moreover, our data indicates that the underlying mechanisms of action involve an IL-17/TNF-alpha synergistic reaction, suppression of Th1 and Th2 responses, and an upregulation of the regulatory cytokines IL-10 and TGF-beta 1. Our results demonstrate that moderate exposure to T. spiralis in the acute phase of DSS-induced colitis improves disease associated inflammation and tissue disruption.

Seroprevalence and associated risk factors of Toxoplasma gondii among Manchu pregnant women in northeastern China.

Toxoplasma gondii is an intracellular protozoan parasite infecting human and almost all warm-blooded animals. It may cause several severe symptoms if pregnant women infection with T. gondii, including misbirth. A cross-sectional study was conducted containing 313 Manchu pregnant women attending antenatal care from 2016 to 2017 in Jilin province, northeastern China, and were assessed by enzyme-link immunosorbent assay and the study utilized univariate analysis to identify the potential risk factors for T. gondii infection. Of the 313 investigated Manchu pregnant women, 51 (16.29%, 95% CI 12.2-20.4) were tested as T. gondii-seropositive, with 47 (15.02%) seropositive for T. gondii IgG antibodies and 8 (2.56%) IgM positive. The seroprevalence of T. gondii in different age groups varied from 13.50% (8.7-18.3) to 23.90% (13.9-41.9). Pregnant raising cat at home has a significantly higher seroprevalence of T. gondii than no cat at home. Pregnant consuming raw/undercooked meat has a significantly higher T. gondii seroprevalence than individuals did not consuming raw/undercooked meat. This is the first study of T. gondii infection seroprevalence in Manchu pregnant women. Risk factors analysis suggested that seroprevalence of T. gondii in investigated Manchu pregnant women was mainly related to consumption of raw/undercooked meat and raising cat at home. The findings will provide key and baseline data for prevention and control of toxoplasmosis among Manchu pregnant women and other people.

Inhibition of GSDMD-mediated pyroptosis triggered by Trichinella spiralis intervention contributes to the alleviation of DSS-induced ulcerative colitis in mice.

BACKGROUND: Inflammatory bowel disease (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), is increasing worldwide. Although there is currently no completely curative treatment, helminthic therapy shows certain therapeutic potential for UC. Many studies have found that Trichinella spiralis (T.s) has a protective effect on UC, but the specific mechanism is still unclear. METHODS: Balb/c mice drank dextran sulfate sodium (DSS) to induce acute colitis and then were treated with T.s. In vitro experiments, the LPS combination with ATP was used to induce the pyroptosis model, followed by intervention with crude protein from T.s (T.s cp). Additionally, the pyroptosis agonist of NSC or the pyroptosis inhibitor vx-765 was added to intervene to explore the role of pyroptosis in DSS-induced acute colitis. The degree of pyroptosis was evaluated by western blot, qPCR and IHC, etc., in vivo and in vitro. RESULTS: T.s intervention significantly inhibited NLRP3 inflammasome activation and GSDMD-mediated pyroptosis by downregulating the expression of pyroptosis-related signatures in vitro (cellular inflammatory model) and in vivo (DSS-induced UC mice model). Furthermore, blockade of GSDMD-mediated pyroptosis by the caspase-1 inhibitor vx-765 has a similar therapeutic effect on DSS-induced UC mice with T.s intervention, thus indicating that T.s intervention alleviated DSS-induced UC in mice by inhibiting GSDMD-mediated pyroptosis. CONCLUSION: This study showed that T.s could alleviate the pathological severity UC via GSDMD-mediated pyroptosis, and it provides new insight into the mechanistic study and application of helminths in treating colitis.

A secreted MIF homologue from Trichinella spiralis binds to and interacts with host monocytes.

Trichinella spiralis is a very successful parasite capable of surviving in many mammal hosts and residing in muscle tissues for long periods, indicating that it must have some effective strategies to escape from or guard against the host immune attack. The functions of MIF have been studied in other parasites and demonstrated to function as a virulence factor aiding in their survival by modulating the host immune response. However, the functions of Trichinella spiralis MIF (TsMIF) have not been addressed. Here, we successfully obtained the purified recombinant TsMIF and anti-TsMIF serum. Our results showed that TsMIF was expressed in all the Trichinella spiralis developmental stages, especially highly expressed in the muscle larvae (ML) and mainly located in stichocytes, midgut, cuticle, muscle cells of ML and around intrauterine embryos of female adults. We also observed TsMIF could be secreted from ML and bind to host monocytes. Next, our data demonstrated that TsMIF not only stimulated the phosphorylation of ERK1/2 and cell proliferation by binding to the host cell surface receptor CD74, but also interacted with a host intracellular protein, Jab1, which is a coactivator of AP-1 transcription. We concluded the secreted TsMIF plays an important role in the interaction between Trichinella spiralis and its host and could be a potential drug or vaccine target molecule against Trichinella spiralis infection.

Trends of cancer incidence among Chinese older adults from 2005 to 2016: A log-linear regression and age-period-cohort analysis.

Background: To study the corresponding strategies for controlling cancer in older adults aged 60 and above in China, a comprehensive assessment of disease burden is required. Therefore, we will introduce the cancer epidemiological characteristics of older adults in China over a recent 12 year period. Methods: The age-period-cohort model was constructed using the cancer incidence data from the Chinese Cancer Registry Annual Report published in 2008-2019. The annual change percentage (APC) was estimated by log-linear regression to reflect the time trend. The data from the GLOBOCAN 2020 database was selected for worldwide comparative analysis. Results: The cancer incidence in older adults aged 60 and above in China showed a decreasing trend (APC = -0.73%, P = 0.009). The urban/rural ratio of cancer incidence increased from 0.94 to 1.07 (t = 3.52, P < 0.05), while the sex ratio (male/female) showed a significant decreasing trend only in rural areas (t = -6.77, P < 0.05), and the ratio decreased from 2.02 to 1.72. The results of the age-period-cohort model showed that the cancer incidence increased with age in both males and females, urban and rural areas. The RR of period effect increased from 2005 to 2010, then decreased from 2010 to 2015, and the downward trend was more obvious. The RR of the later-born cohort was lower than that of the earlier-born cohort in rural areas. Lung, gastric, colorectal, esophageal, liver, and breast cancers were common cancers in Chinese older adults. Lung cancer incidence ranked first in males, and it decreased with time in the 75-79 and 80-84 age groups (APC75 - 79 = -1.10%, APC80 - 84 = -0.88%, all P < 0.05). Breast cancer incidence ranked first among female in the 60-64 age group and showed an increasing trend (APC60 - 64 = 1.52%, P < 0.05). Conclusions: The cancer incidence in Chinese older adults aged 60 and above showed a decreasing trend, but it was still at a relatively high level. The key targets of prevention and treatment should be males, urban areas, younger people, older adults aged 60-69, lung, gastrointestinal, and breast cancers in the future.

Modeling manufacturing resources based on manufacturability features.

Manufacturability evaluation is an effective way to shorten the development period, optimize manufacturing processes, and reduce product costs. The manufacturability of a product depends on the processing ability of specific manufacturing resources. The development of a manufacturing resources model serves as the foundation for manufacturability evaluation. To better utilize the information on manufacturing resources, this study adopted a hybrid approach by integrating the fuzzy c-means clustering algorithm and the genetic algorithm to group manufacturing resources based on manufacturing and geometric features. The information model of manufacturing resources was built by using the object-oriented method. Subsequently, the framework to evaluate manufacturing capability based on manufacturing resources was defined. Further, an application sample was exploited and its results were analyzed. The results of the subgroup showed that the hybrid algorithm was reliable and valid and helped improve the overall performance of the company chosen for this study. The proposed approach enhanced feasibility in decision-making and facilitated the management to make more informed decisions.

Virulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway.

BACKGROUND: Toxoplasma gondii is known as the most successful parasite, which can regulate the host immune response through a variety of ways to achieve immune escape. We previously reported that a novel gene wx2 of T. gondii may be a virulence-related molecule. The objective of this study was to explore the mechanism of wx2 regulating host immune response. METHODS: The wx2 knockout strain (RHwx2-/- strain) and complementary strain (RHwx2+/+ strain) were constructed by the CRISPR/Cas9 technique, and the virulence of the wx2 gene was detected and changes in pyroptosis-related molecules were observed. RESULTS: Compared with the wild RH and RHwx2+/+ strain groups, the survival time for mice infected with the RHwx2-/- strain was prolonged to a certain extent. The mRNA levels of pyroptosis-related molecules of caspase-1, NLRP3, and GSDMD and et al. in mouse lymphocytes in vivo and RAW267.4 cells in vitro infected with RHwx2-/- strain increased to different degrees, compared with infected with wild RH strain and RHwx2+/+ strain. As with the mRNA level, the protein level of caspase-1, caspase-1 p20, IL-1ß, NLRP3, GSDMD-FL, GSDMD-N, and phosphorylation level of NF-κB (p65) were also significantly increased. These data suggest that wx2 may regulate the host immune response through the pyroptosis pathway. In infected RAW264.7 cells at 48 h post-infection, the levels of Th1-type cytokines of IFN-γ, Th2-type cytokines such as IL-13, Th17-type cytokine of IL-17 in cells infected with RHwx2-/- were significantly higher than those of RH and RHwx2+/+ strains, suggesting that the wx2 may inhibit the host's immune response. CONCLUSION: wx2 is a virulence related gene of T. gondii, and may be involved in host immune regulation by inhibiting the pyroptosis pathway.

Epitope vaccine design for Toxoplasma gondii based on a genome-wide database of membrane proteins.

BACKGROUND: There is presently no effective and safe vaccine for Toxoplasma gondii for humans. The study described here was designed to search for a novel group of optimal B cell and T cell epitopes from Toxoplasma membrane proteins using genome-wide comprehensive screening. METHODS: The amino acid sequences of membrane proteins of T. gondii were obtained from the UniProt database. The ABCPred and BepiPred servers were employed to predict the linear B cell epitopes. The Immune Epitope Database (IEDB) online service was utilized to forecast T cell epitopes within T. gondii membrane proteins that bind to human leukocyte antigen (HLA) class I (HLA-I) or HLA-II molecules. RESULTS: From the 314 membrane proteins of T. gondii, a total of 14 linear B cell epitopes embedded in 12 membrane proteins were identified. Eight epitopes for major histocompatibility complex (MHC) class I (MHC-I) molecules and 18 epitopes for MHC-II molecules were ultimately selected, for which world population coverage percentiles were 71.94% and 99.76%, respectively. The top rated combinations of linear B cell epitopes and T cell epitopes covering both BALB/c mice and a majority of the human population were identified for the development of a protective vaccine. CONCLUSIONS: The ultimate vaccine construct described here, which comprises B cells, MHC-I and MHC-II epitopes, might protect individuals against T. gondii infection by inducing humoral and cellular immune responses.

A Novel wx2 Gene of Toxoplasma gondii Inhibits the Parasitic Invasion and Proliferation in vitro and Attenuates Virulence in vivo via Immune Response Modulation.

Toxoplasma gondii (T. gondii) is an obligate intracellular apicomplexan protozoan that can parasitize most warm-blooded animals and cause severe diseases in immunocompromised individuals or fetal abnormalities in pregnant woman. The treatment of toxoplamosis has been limited by effective drugs. Our previous work indicated that the novel gene wx2 of T. gondii may serve as a vaccine antigen candidate. To further investigate the molecular functions of wx2 in highly virulent T. gondii (RH strain), a wx2 gene deletion mutant RH strain (KO-wx2) was established using CRISPR-Cas9. The phynotype of KO-wx2 was analyzed by plaque, invasion, and replication assays in vitro as well as in vivo virulence assays. The results indicated that the targeted deletion of the wx2 gene significantly inhibited in vitro parasite growth and replication in the host cells as well as attenuated parasite virulence in the mouse model. Notably, the percentage of pro-inflammatory factors of interferon gamma (IFN-γ) and interlukin-17A (IL-17A) and anti-inflammatory factor of interlukin-10 (IL-10) in the lymph nodes were upregulated in mice infected with the KO-wx2 strain. Our data suggested that the wx2 gene plays an important role in the process of the parasite's life cycle and virulence in mice. In addition, it also plays an important role in the host's immunity reaction, mainly via Th1 and Th17 cellular immunity, not Th2.

Corrigendum: A Novel wx2 Gene of Toxoplasma gondii Inhibits the Parasitic Invasion and Proliferation in vitro and Attenuates Virulence in vivo via Immune Response Modulation.

[This corrects the article DOI: 10.3389/fmicb.2020.00399.].

Matrix Gla Protein Promotes the Bone Formation by Up-Regulating Wnt/ß-Catenin Signaling Pathway.

Objective: Studies suggest that matrix Gla protein (MGP) is associated with osteoporosis. However, the precise mechanism through which MGP regulates bone metabolism is not fully understood. The purpose of this study was to clarify the role of MGP in bone metabolism. Methods: The MGP gene in MG63 cell line was knocked down using shRNA. Cell Counting Kit-8 assay was used to detect the proliferation of MG63 cells. Moreover, the differentiation and mineralization of MG63 cells were measured through alkaline phosphatase staining and Alizarin Red S staining. Western blotting and quantitative reverse transcription-polymerase chain reaction were conducted to detect the protein and mRNA levels of components of the Wnt/ß-catenin signaling pathway, such as Wnt3a, ß-catenin, and Runx2. Transgenic (MGP+) mice were used to detect the effects of MGP in vivo. Results: The Cell Counting Kit-8 assay suggested that upregulated MGP could promote the proliferation of MG63 cells, whereas its downregulation inhibited proliferation. The alkaline phosphatase assay and Alizarin Red S staining showed that overexpressed MGP led to prominently upregulated differentiation and mineralization of MG63 cells. Conversely, knockdown of MGP decreased the levels of differentiation and mineralization. Western blotting and quantitative reverse transcription-polymerase chain reaction showed that overexpression of MGP upregulated Wnt3a, ß-catenin, and Runx2. In contrast, knocking down MGP reduced their transcriptional levels. In vivo, overexpression of MGP inhibited the decrease in bone mineral density induced via ovariectomy in the femur, and significantly prevented bone volume fraction, trabecular number, BV/TV, and TbTh to decrease. In addition, it increased the levels of estradiol in sera. Conclusion: The findings of this study suggest that the promotion of osteoblast proliferation, differentiation, and mineralization by MGP may be a mechanism to prevent osteoporosis. Furthermore, the results show that MGP promoted the osteogenic effects via the Wnt/ß-catenin signaling pathway.

Effect of fertilizer and water content on N2O emission from three plantation soils in south China.

The effects of fertilizers and water content on N2O emission were studied using the three most typical plantation soils. Soil incubations were performed and fertilization and water content treatments were designed. At 25% of saturated water content(SWC), N2O emissions from the soil treated with urea, KNO3, (NH4)2 SO4 and KH2 PO4 were compared at application rates of 0, 100, 200, 300 and 500 kg/hm2. At 80% of SWC, similar experiments were carried out but at only one application rate(500 kg/hm2). N2O emissions at various water contents(20%, 35%, 50%, 65%, 80% and 100% of SWC) were studied. At low water content(25% of SWC), neither nitrogen nor phosphorus(or potassium) fertilizers led to a high level of N2O emission, which generally ranged from 2.03 to 29.02 microg/(m2 x h). However, at high water content(80% SWC), the fertilizers resulted in much greater N2O emission irregardless of soil tested. The highest N2O emission rates after 24 h of water addition were 1233 microg/(m2 x h) for S. superba soil, 1507 microg/(m2 x h) for P. elliottii soil and 1869 microg/ (m2 x h) for A. mangium soil respectively. N2O emission from soils treated with urea, (NH4)2 SO4 and KH2 PO4 immediately dropped to a low level but steadily increased to a very high level for the soil treated with KNO3. High NO3- content was a basis of high level of N2O emission. N2O emission rates from soils peaked shortly after flooding, rapidly dropping to a very low level in soil from non-legume plantations, but lasting for a relatively long period in soil from legume plantations. When soil water content increased equaling to or higher than 65%, the accumulated N2O emission over a period of 13 d ranged from 20.21-29.78 mg/m2 for S. superba, 30.57-70.12 mg/m2 for P. elliottii and 300.89-430.51 mg/m2 for A. mangium. The critical water content was 50% of SWC, above which a high level of N2O emission could be expected, and below which very little N2O emissions were detected. The results suggest that, at low water content (< 50% of SWC), the fertilization practice is safe with regard to N2O emissions, but at high water content (> 50% of SWC), nitrogen fertilizer in the form of nitrate could yield a 100-fold increase in N2O emissions. Legume plantations like A. mangium should be avoided in low lands which could easily suffer from flooding or poor drainage.