Spirometra erinaceieuropaei casein kinase I (SeCKI) was analyzed using bioinformatical methods to predict its structure and function based on the deduced amino acid sequence from full length cDNA sequence of SeCKI gene with online sites and software programs. The longest open reading frame contains 448 amino acids, 50 kDa and theoretical pI of 4.73, with a complete tubulin domain, a SMART tubulin_C domain and a low complexity region. SeCKI has no signal sequence and no transmembrane domain, but is predicted to be located extracellularly. The secondary structure of SeCKI contains 12 α-helixes, 11 β-strands and 22 coils. SeCKI had 19 potential antigenic epitopes and 25 HLA-I restricted epitopes. Based on phylogenetic analysis of SeCKI sequence, S. erinaceieuropaei has the closest evolutionary status with Hymenolepis microstoma. Information from this study could provide important insights into the identification of diagnostic antigens and molecular targets of antisparganum drugs.

We have previously reported that the recombinant T. spiralis aminopeptidase (rTsAP) could induce a partial protective immunity against T. spiralis infection in mice. The aim of this study was to predict the structures and functions of TsAP protein by using the full length cDNA sequence of TsAP gene. TsAP sequence was 1515 bp length with a 1515 bp biggest ORF encoding 504-amino acid protein. The molecular weight and isoelectric point of TsAP were 54.7 kDa and 6.69, respectively. TsAP structure domains contained a Peptidase_M17_N and a Peptidase_M17 domain, which has the function of catalysis of the hydrolysis of N-terminal amino acid residues. TsAP had no signal peptide site and transmembrane domain, and located in cytoplasm. The secondary structure of TsAP contained 16 α-helix, 14 β-strand and 29 coils. The TsAP had 11 and 21 potential antigenic epitopes of T cell and B cell, respectively. Based on the phylogenetic analyses of TsAP, T. spiralis have the closest relationship with Plasmodium falciparum. TsAP was a kind of proteolytic enzyme with a variety of biological functions and its antigenic epitopes could provide important insights on the diagnostic antigens and target molecular of anti-Trichinella drugs

Previous studies showed that crude antigens from Trichinella spiralis adult worms (AW) can be recognized by mouse infection sera at 8 days post infection. The aim of this study was to identify the early diagnostic antigenic bands in soluble proteins from T. spiralis AW by Western blot using early infection sera. The affecting factors of adult recovery were firstly observed in this study, and the results showed that the maximum number of adults was collected from small intestine when the female BALB/c mice were orally infected with 4000 ML and sacrificed at 3 days post infection. The results of Western blot analysis showed that seven protein bands (31, 35.1, 39, 40.6, 41.9, 47 and 50.6 kDa) could be recognized by early infection sera as early as at 8-10 days post infection, and were strongly reacted with mouse infection sera at 11-12 days post infection. Our results suggested that the seven protein bands of T. spiralis AW soluble proteins might be the early expressed antigens during the intestinal stage of Trichinella infection and therefore have potential value for the early diagnosis of trichinellosis.

Objective: To analyze the characteristics of super-antigen (SAg) of group A Streptococcus pyogenes (GAS), isolated from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017. Methods: Throat swab specimens from patients with scarlet fever or pharyngeal infections were collected and tested for GAS. Eleven currently known SAg genes including SpeA, speC, speG, speH, speI, speJ, speK, speL, speM, smeZ and ssa were tested by real-time PCR while M protein genes (emm genes) were amplified and sequenced by PCR. Results: A total of 377 GAS were isolated from 6 801 throat swab specimens, with the positive rate as 5.5%. There were obvious changes noticed among speC, speG, speH and speK in three years. A total of 45 SAg genes profiles were observed, according to the SAgs inclusion. There were significant differences appeared in the frequencies among two of the highest SAg genes profiles between emm1 and emm12 strains (χ(2)=38.196, P<0.001; χ(2)=72.310, P<0.001). There also appeared significant differences in the frequencies of speA, speH, speI and speJ between emm1 and emm12 strains (χ(2)=146.154, P<0.001; χ(2)=52.31, P<0.001; χ(2)=58.43, P<0.001; χ(2)=144.70, P<0.001). Conclusions: Obvious changes were noticed among SAg genes including speC, speG, speH and speK from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017. SAg genes including speA, speH, speI and speJ appeared to be associated with the emm 1 and emm 12 strains. More kinds of SAg genes profiles were isolated form GAS but with no significant differences seen in the main SAg genes profiles, during the epidemic period.
Antigens, Bacterial/genetics* ; Bacterial Outer Membrane Proteins ; Bacterial Proteins ; Beijing/epidemiology* ; China/epidemiology* ; Exotoxins ; Female ; Humans ; Membrane Proteins ; Pharyngitis/microbiology* ; Pharynx/microbiology* ; Pregnancy ; Pregnancy Complications, Infectious/microbiology* ; Real-Time Polymerase Chain Reaction ; Scarlet Fever/microbiology* ; Streptococcal Infections ; Streptococcus pyogenes/isolation & purification* ; Superantigens/genetics*

Spirometra larvae are etiological agents of human sparganosis. However, the systematics of spirometrid cestodes has long been controversial. In order to determine the current knowledge on the evolution and genetic structure of Spirometra, an exhaustive population diversity analysis of spirometrid cestodes using the mitochondrial gene: cytochrome c oxidase subunit 1 (cox1) was performed. All publicly available cox1 sequences available in the GenBank and 127 new sequencing genes from China were used as the dataset. The haplotype identify, network, genetic differentiation and phylogenetic analysis were conducted successively. A total of 488 sequences from 20 host species, representing four spirometrid tapeworms (S. decipiens, S. ranarum, S. erinaceieuropaei and Sparganum proliferum) and several unclassified American and African isolates from 113 geographical locations in 17 countries, identified 45 haplotypes. The genetic analysis revealed that there are four clades of spirometrid cestodes: Clade 1 (Brazil + USA) and Clade 2 (Argentina + Venezuela) included isolates from America, Clade 3 contained African isolates and one Korean sample, and the remainders from Asia and Australia belonged to Clade 4; unclassified Spirometra from America and Africa should be considered the separate species within the genus; and the taxonomy of two Korea isolates (S. erinaceieuropaei KJ599680 and S. decipiens KJ599679) was still ambiguous and needs to be further identified. In addition, the demographical analyses supported population expansion for the total spirometrid population. In summary, four lineages were found in the spirometrid tapeworm, and further investigation with deeper sampling is needed to elucidate the population structure.

Objective: To understand the awareness of hepatic disease related knowledge among hepatic physicians in poverty-stricken counties in China, assess the effectiveness of training and provide a reference for the training in the future. Methods: The training was conducted in 90 clinical hepatic physicians selected from county hospitals in poverty-stricken counties (or cities) in Shanxi and Shaanxi provinces. An examination was conducted before the training, immediately after the training and at 5(th) month after the training, respectively. One-way analysis of variance and χ(2) test were conducted to evaluate the score and the correct rate. Results: The knowledge score was (42.96±14.02) before the training, (62.86±13.28) immediately after the training and (59.03±17.92) at 5(t)h month after the training, and the differences were significant. After the training, the awareness of all aspects of related knowledge was improved, the difference was significant compared to knowledge score before training, and at 5(th) month after the training, the difference was still significant. Conclusion: After the training, the awareness of liver disease related knowledge of clinical hepatic physicians in poverty-stricken counties (cities) in Shanxi and Shaanxi provinces was improved, and the improvement could be maintained for nearly half a year.
Awareness ; China/epidemiology* ; Clinical Competence ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Liver Diseases/therapy* ; Physicians ; Poverty Areas ; Program Evaluation ; Staff Development/methods* ; Surveys and Questionnaires

In previous studies, a Trichinella spiralis serine protease (TsSP) was identified in excretion/secretion (ES) products from intestinal infective L1 larvae (IIL1) using immunoproteomics. The complete cDNA sequence of TsSP gene was 1372 bp, which encoded 429 amino acids with 47.55 kDa. The TsSP was transcribed and expressed at all T. spiralis life cycle phases, as well as mainly located at the cuticle and stichosome of the parasitic nematode. Recombinant TsSP bind to intestinal epithelial cells (IEC) and promoted larva invasion, however, its exact function in invasion, development and reproduction are still unknown. The aim of this study was to confirm the biological function of TsSP during T. spiralis invasion and growth using RNA interference (RNAi) technology. The results showed that on 1 day after electroporation using 2.5 µM siRNA156, TsSP mRNA and protein expression of muscle larvae (ML) was suppressed by 48.35 and 59.98%, respectively. Meanwhile, silencing of TsSP gene by RNAi resulted in a 61.38% decrease of serine protease activity of ML ES proteins, and a significant reduction of the in vitro and in vivo invasive capacity of IIL1 to intrude into the IEC monolayer and intestinal mucosa. When mice were infected with siRNA 156-transfected larvae, adult worm and muscle larva burdens were decreased by 58.85 and 60.48%, respectively. Moreover, intestinal worm growth and female fecundity were evidently inhibited after TsSP gene was knockdown, it was demonstrated that intestinal adults became smaller and the in vitro newborn larval yield of females obviously declined compared with the control siRNA group. The results indicated that knockdown of TsSP gene by RNAi significantly reduced the TsSP expression and enzymatic activity, impaired larvae intrusion and growth, and lowered the female reproductive capacity, further verified that TsSP might participate in diverse processes of T. spiralis life cycle, it will be a new prospective candidate molecular target of anti-Trichinella vaccines.

Objective: To investigate the association between alcohol consumption and lung cancer risk in Chinese males. Methods: Information on alcohol consumption and outcomes were collected on a biennial basis among males in Kailuan Cohort (2006-2015). In addition, electronic databases of hospitals affiliated to Kailuan Community, Insurance Systems of Kailuan Community and Tangshan were also used for supplementary information retrieval. Cox proportional hazards regression models were used to evaluate the hazard ratio (HR) and 95%CI of baseline frequency and type of alcohol consumption associated with lung cancer risk in males. Non-drinkers were used as control group. Results: A total of 101 751 males were included and 913 new lung cancer cases were identified in the Kailuan male cohort study, with a total follow-up time of 808 146.56 person-years and a median follow-up time of 8.88 years by 31 December 2015. After adjusting for potential confounding factors, the HR of former drinkers, occasional drinkers (<1/day) and drinkers (≥1/day) were 1.30 (95%CI: 0.90-1.88), 0.80 (95%CI: 0.64-1.01) and 1.04 (95%CI: 0.85-1.27), respectively, compared with non-drinkers. In addition, drinking beer/red wine (HR=0.91, 95%CI: 0.69-1.20) and white wine (HR=0.99, 95%CI: 0.83-1.19) showed no significant association with lung cancer. The results were similar when stratified analysis were conducted. Conclusion: Our study results don't support the hypothesis that alcohol consumption is significantly associated with the risk of lung cancer in males.
Adult ; Alcohol Drinking/epidemiology* ; China/epidemiology* ; Cohort Studies ; Humans ; Lung Neoplasms/epidemiology* ; Male ; Middle Aged ; Proportional Hazards Models ; Prospective Studies ; Risk Factors

Trichinellosis is an important zoonotic parasitic disease worldwide and is principally caused by ingesting animal meat containing Trichinella infective larvae. Aspartyl aminopeptidase is an intracytoplasmic metalloproteinase that specifically hydrolyzes the N-terminus of polypeptides free of acidic amino acids (aspartic acid and glutamate), and plays an important role in the metabolism, growth and development of organisms. In this study, a novel T. spiralis aspartyl aminopeptidase (TsAAP) was cloned and expressed, and its biological properties and roles in worm growth and development were investigated. The results revealed that TsAAP transcription and expression in diverse T. spiralis stages were detected by RT-PCR and Western blotting, and primarily localized at cuticle, stichosome and intrauterine embryos of this nematode by immunofluorescence test. rTsAAP has the enzymatic activity of native AAP to hydrolyze the substrate H-Glu-pNA. There was a specific binding between rTsAAP and murine erythrocyte, and the binding site was localized in erythrocyte membrane proteins. Silencing of TsAAP gene by specific dsRNA significantly reduced the TsAAP expression, enzymatic activity, intestinal worm burdens and female fecundity. The results demonstrated that TsAAP participates in the growth, development and fecundity of T. spiralis and it might be a potential target molecule for anti-Trichinella vaccines.

A T. spiralis serine protease 1.2 (TsSP1.2) was identified in the muscle larvae (ML) and intestinal larvae surface/excretory–secretory (ES) proteins by immunoproteomics. The aim of this study was to determine the TsSP1.2 function in the process of T. spiralis intrusion, growth and reproduction by using RNA interference (RNAi). RNAi was used to silence the expression of TsSP1.2 mRNA and protein in the nematode. On 2 days after the ML were electroporated with 2 µM of TsSP1.2-specific siRNA 534, TsSP1.2 mRNA and protein expression declined in 56.44 and 84.48%, respectively, compared with untreated ML. Although TsSP1.2 silencing did not impair worm viability, larval intrusion of intestinal epithelium cells (IEC) was suppressed by 57.18% (P < 0.01) and the suppression was siRNA-dose dependent (r = 0.976). Infection of mice with siRNA 534 transfected ML produced a 57.16% reduction of enteral adult burden and 71.46% reduction of muscle larva burden (P < 0.05). Moreover, silencing of TsSP1.2 gene in ML resulted in worm development impediment and reduction of female fertility. The results showed that silencing of TsSP1.2 by RNAi inhibited larval intrusion and development, and reduced female fecundity. TsSP1.2 plays a crucial role for worm invasion and development in T. spiralis life cycle, and is a potential vaccine/drug target against Trichinella infection.