Exosomal circ_0032704 confers sorafenib resistance to hepatocellular carcinoma and contributes to cancer malignant progression by modulating the miR-514a-3p/PD-L1 pathway.

Purpose: This study aims to explore the role of circ_0032704 in sorafenib-resistant hepatocellular carcinoma (HCC). Methods: The expression of circ_0032704, miR-514a-3p, and programmed death-ligand 1 (PD-L1) mRNA was detected by quantitative real-time PCR (qPCR). The expression of multidrug resistant-related proteins, migration/invasion-related proteins, exosome-related proteins, and PD-L1 protein was detected by western blot. Cell viability was detected by CCK-8 assay. Cell proliferation, migration, and invasion were assessed by EdU assay, wound healing assay, and transwell assay. The binding between miR-514a-3p and circ_0032704 or PD-L1 was verified by RIP assay, pull-down assay, and dual-luciferase reporter assay. Cell- or serum-derived exosomes were isolated and identified by TEM and NTA. Xenograft models were established to determine the effect of circ_0032704 on drug resistance in vivo. Results: Circ_0032704 was overexpressed in sorafenib-resistant HCC tissues and cells. Circ_0032704 knockdown reduced sorafenib resistance in HCC cells and inhibited cell proliferation, migration, and invasion of sorafenib-resistant HCC cells, while these effects were reversed by PD-L1 overexpression. We found that circ_0032704 positively regulated PD-L1 expression via targeting miR-514a-3p. Exosomes with circ_0032704 inhibition reduced sorafenib resistance in HCC cells and inhibited cell proliferation, migration, and invasion of sorafenib-resistant HCC cells. Exosomes with circ_0032704 inhibition also inhibited tumor growth in vivo. The expression of circ_0032704 in exosomes was stable and possessed diagnostic value. Conclusion: Circ_0032704 enhanced sorafenib resistance in HCC and promoted the malignant development of sorafenib-resistant HCC. Circ_0032704 could be transported by exosomes, and exosomal circ_0032704 had diagnostic value.

Integrated Pharmaco-Bioinformatics Approaches and Experimental Verification To Explore the Effect of Britanin on Nonalcoholic Fatty Liver Disease.

Nonalcoholic fatty liver disease (NAFLD) is a prevalent global liver disorder, posing substantial health risks. Britanin, a bioactive sesquiterpene lactone extracted from Inula japonica, has demonstrated antidiabetic, hypolipidemic, and hepatoprotective attributes. Nonetheless, the precise impact of Britanin on NAFLD and the intricate biological mechanisms underpinning this interaction remain unexplored. We integrated computer-aided methods to unearth shared biological targets and signaling pathways associated with both Britanin and NAFLD. A network was constructed by compiling putative targets associated with Britanin and NAFLD, followed by a stringent screening of key targets and mechanisms through protein-protein interaction analysis along with GO and KEGG pathway enrichment analyses. Molecular docking was integrated as an evaluation tool, culminating in the identification of HO-1 as the pivotal therapeutic target, showcasing a satisfactory binding affinity. The primary mechanism was ascribed to biological processes and pathways linked to oxidative stress, as evidenced by the outcomes of enrichment analyses. Of these, the AMPK/SREBP1c pathway assumed centrality in this mechanism. Furthermore, in vivo experiments substantiated that Britanin effectively curtailed NAFLD development by ameliorating liver injury, modulating hyperlipidemia and hepatic lipid accumulation, and alleviating oxidative stress and apoptosis. In summary, this study demonstrates the potential of Britanin as a promising therapeutic drug against NAFLD.

Associated factors of depression in primiparas with hypothyroidism during pregnancy.

This cross-sectional study aimed to explore the associated factors of depression in primiparas with hypothyroidism during pregnancy. The research subjects were 200 primiparas with hypothyroidism during pregnancy who were admitted to our hospital between December 2016 and December 2019. Self-rating depression scale scores were used to evaluate the depression, and the incidence of depression were examined. The data from all the subjects were collected to compare the differences between primiparas with hypothyroidism during pregnancy with and without depression. A logistic regression equation was used to analyze the influencing factors of depression in these patients. Of the 200 primiparas who took part in this study, 27 suffered from depression, accounting for 13.50%. There were differences in age, education level, economic income, sleep quality, and conjugal relations between the depressed and the nondepressed participants. When the above factors were included in the logistic regression equation, it was found that the odds ratio values for these factors were all >1, which indicated that they had an influence on maternal depression in primiparas with hypothyroidism during pregnancy. This study demonstrated that pregnancy-associated hypothyroidism in primiparas is affected by age, education level, economic income, sleep quality, and conjugal relations, all of which increase the incidence of depression. Relevant preventive measures should be provided in clinical practice to avoid the occurrence of depression.

Effect of Internet + continuous midwifery service model on psychological mood and pregnancy outcomes for women with high-risk pregnancies.

BACKGROUND: There are many drawbacks to the traditional midwifery service management model, which can no longer meet the needs of the new era. The Internet + continuous midwifery service management model extends maternal management from prenatal to postpartum, in-hospital to out-of-hospital, and offline to online, thereby improving maternal and infant outcomes. Applying the Internet + continuous midwifery service management model to manage women with high-risk pregnancies (HRP) can improve their psycho-emotional opinion and, in turn, minimize the risk of adverse maternal and/or fetal outcomes. AIM: To explore the effectiveness of a midwife-led Internet + continuous midwifery service model for women with HRP. METHODS: We retrospectively analyzed the clinical data of 439 women with HRP who underwent prenatal examination and delivered at Shanghai Sixth People's Hospital (affiliated to the Shanghai Jiao Tong University School of Medicine) from April to December 2022. Among them, 239 pregnant women underwent routine obstetric management, and 200 pregnant women underwent Internet + continuous midwifery service mode management. We used the State-Trait Anxiety Inventory, Edinburgh Postnatal Depression Scale, and analysis of delivery outcomes to compare psychological mood and the incidence of adverse delivery outcomes between the two groups. RESULTS: The data showed that in early pregnancy, the anxiety and depression levels of the two groups were similar; the levels gradually decreased as pregnancy progressed, and the decrease in the continuous group was more significant [31.00 (29.00, 34.00) vs 34.00 (32.00, 37.00), 8.00 (6.00, 9.00) vs 12.00 (10.00, 13.00), P < 0.05]. The maternal self-efficacy level and strategy for weight gain management were better in the continuous group than in the traditional group, and the effective rate of midwifery service intervention in the continuous group was significantly higher than in the control group [267.50 (242.25, 284.75) vs 256.00 (233.00, 278.00), 74.00 (69.00, 78.00) vs 71.00 (63.00, 78.00), P < 0.05]. The incidence of adverse delivery outcomes in pregnant women and newborns and fear of maternal childbirth were lower in the continuous group than in the traditional group, and nursing satisfaction was higher [10.50% vs 18.83%, 8.50% vs 15.90%, 24.00% vs 42.68%, 89.50% vs 76.15%, P < 0.05]. CONCLUSION: The Internet + continuous midwifery service model promotes innovation through integration and is of great significance for improving and promoting maternal and child health in HRP.

Regression analysis of bleeding factors and the correlation between FPE and NLR after mechanical thrombectomy for acute anterior circulation large vessel occlusion.

Acute anterior circulation large vessel occlusion refers to a blockage in the blood flow of one of the major blood vessels in the anterior (front) part of the brain. Acute anterior circulation large vessel occlusion can lead to a range of symptoms, including sudden headache, difficulty speaking or understanding speech, weakness or numbness on one side of the body and loss of vision in one eye. According to relevant data, mechanical thrombectomy in the treatment of the large vessel recanalization rate can reach 70%. However, hemorrhage is one of the serious complications after mechanical thrombectomy, and it is the main factor leading to the deterioration of neurological function and death of patients after large blood vessels. Therefore, the risk factors of bleeding in patients were analyzed before surgery, and effective prevention during and after surgery had positive significance for patients undergoing mechanical thrombectomy. This study uses regression analysis to analyze the correlation between bleeding factors and FPE and NLR after mechanical thrombectomy for acute anterior circulation large vessel occlusion. We retrospectively analyzed 81 patients with acute anterior circulation large vessel occlusion who underwent mechanical embolization in our hospital from September 2019 to January 2022 and separated them into a bleeding group (46 patients) and a non-bleeding group (35 patients) based on the presence or absence of bleeding after surgery.

The Prevalence and Influencing Factors of Postpartum Depression Between Primiparous and Secundiparous.

ABSTRACT: Since the implementation of "two-child policy" in China, the prevalence and influencing factors of postpartum depression (PPD) between women at first childbirth and second childbirth remains unclear. We aimed to compare the prevalence and the influencing factors of PPD between the two groups. Postpartum women were recruited at their 6-week postpartum checkups and invited to complete questionnaires. The prevalence rate of PPD was 42.1%. There was no statistical difference found regarding the prevalence of PPD and the total score on the Edinburgh Postnatal Depression Scale between the two groups. The median of the item "self-injury" was significantly higher among women at first childbirth. Women with gestational diabetes in pregnancy, antenatal anxiety and depression, abortion, having babies getting sick occasionally, and deteriorated relationships with partner in pregnancy had much higher risks for PPD. Both primiparous and secundiparous had a high prevalence of PPD. It provides scientific evidence for clinical preventive and nursing care for PPD.

The prevalence of bovine mastitis-associated Staphylococcus aureus in China and its antimicrobial resistance rate: A meta-analysis.

In this study, to optimize the Staphylococcus aureus control program, a meta-analysis was conducted to investigate the epidemiology and antimicrobial resistance (AMR) profile of S. aureus-associated bovine mastitis in China from 2000 to 2020. A total of 33 publications from PubMed, Google Scholar, and China National Knowledge Infrastructure (CNKI) database were included in our research, among which nine publications included the AMR test. The pooled prevalence of S. aureus was 36.23%, and subgroup analysis revealed that the prevalence dropped from 2000-2010 to 2011-2020, which shows that China is on the right track. The pooled AMR rate indicate isolates were most resistant to ß-lactams (50.68%), followed by quinolones (36.23%), macrolides (34.08%), sulfonamides (32.25%), tetracyclines (27.83%), aminoglycosides (26.44%), lincosamides (23.39%), and amphenicol (10.33%). Both the pooled prevalence and AMR of S. aureus in China are higher than those in Western countries, such as Germany, Belgium, Ireland, and the United States-countries with a long animal husbandry history and good management. Thus, there is still room to improve the treatment of S. aureus-associated bovine mastitis in China.

Molecular Detection and Multilocus Genotyping of Giardia duodenalis in Pigs in Fujian Province, Southeastern China.

Giardia duodenalis, an intestinal parasite, is widely distributed in humans and various animals, such as pigs, cattle and cats. The clinical symptoms of giardiasis are characterized as including abdominal pain, acute or chronic diarrhea, and bloating and weight loss in humans and animals, leading to public and veterinary health problems worldwide. However, the prevalence and genotypes of G. duodenalis in pigs in Fujian Province, southeastern China, have not been reported. In the present study, 725 fecal samples were collected from six cities (Fuqing, Putian, Nanping, Longyan, Sanming, Zhangzhou) in Fujian Province and analyzed for G. duodenalis prevalence and genotypes using nested PCR targeting the beta-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. The results shown that total occurrence rate of G. duodenalis was 26.9% (195/725) in pigs, with significant differences in the prevalence among different regions (χ2 = 86.508, p < 0.05) and groups (χ2 = 12.748, p < 0.05). 195, 11 and 6 samples were detected at the bg, tpi and gdh loci, respectively. Each one belonged to a subtype of assemblage E and was analyzed using sequences obtained in this study. Based on phylogenetic analyses of sequences from the three genetic loci, only one MLG E1 was found. The results indicated that pigs may present a potential zoonotic risk of spreading G. duodenalis infection from animals to humans in this area. The findings of the present study also provide basic data for the prevention and control of G. duodenalis infection in pigs and humans in China.

Molecular Characteristics and Pathogenicity of a Novel Recombinant Porcine Reproductive and Respiratory Syndrome Virus Strain from NADC30-, NADC34-, and JXA1-Like Strains That Emerged in China.

Recently, the emergence of a NADC34-like porcine reproductive and respiratory syndrome virus (PRRSV), which causes a large number of abortions in swine herds, has raised great concern in China. In this study, a PRRSV variant strain, PRRSV/CN/FJGD01/2021, evolved from recombination between NADC30-like, NADC34-like, and JXA1-like viruses was isolated in Fujian province in 2021, and its pathogenicity in piglets was examined. Animal experiments demonstrated that PRRSV/CN/FJGD01/2021 infection could induce 100% morbidity and cause higher viremia, a persistently higher fever (>40°C for 14 consecutive days), significant weight loss, and severe histopathological lung lesions compared to the NADC30-like FJZ03 strain and NADC34-like FJ0908 strain in piglets. The PRRSV/CN/FJGD01/2021 strain displayed higher pathogenicity than the FJZ03 and FJ0908 strains, but lower pathogenicity than the Chinese highly pathogenic (HP)-PRRSVs in piglets. Moreover, the Ingelvac PRRS modified live vaccine (MLV) provides incomplete cross-protection against heterologous PRRSV/CN/FJGD01/2021 in piglets. Our findings contribute to the understanding of the current epidemic situation of NADC34-like PRRSV in China. IMPORTANCE The pathogenicity of NADC34-like PRRSV has broad variations in virulence. Importantly, NADC34-like PRRSV has undergone complex recombination with local strains since it first emerged in 2017 in China. However, the pathogenicity of the recombinant NADC34-like virus was rarely experimentally evaluated in pigs. In this study, a novel PRRSV strain, PRRSV/CN/FJGD01/2021, was isolated from sows enduring a high-abortion-rate (20%) period in China in 2021. Notably, phylogenetic and recombination analyses revealed that PRRSV/CN/FJGD01/2021 is a recombinant virus from NADC30-, NADC34-, and JXA1-like isolates. PRRSV/CN/FJGD01/2021 was shown to cause higher virus load, persistent fever, significant weight loss, moderate respiratory clinical signs, and severe histopathological lung lesions in piglets. PRRSV/CN/FJGD01/2021 exhibited higher pathogenicity than NADC30-like FJZ03 and NADC34-like FJ0908, but lower than Chinese HP-PRRSVs for piglets. These data indicated that PRRSV/CN/FJGD01/2021 has intermediate virulence for piglets. Furthermore, the Ingelvac PRRS MLV could partly provide protective efficacy against PRRSV/CN/FJGD01/2021 challenge in piglets.

Molecular identification of Colpodella sp. of South China tiger Panthera tigris amoyensis (Hilzheimer) in the Meihua Mountains, Fujian, China.

A three-year-old male South China tiger died in the tiger enclosure of the China Tiger Park in the Meihua Mountains on December 2018 after being bitten by a tick. This tiger presented clinical symptoms like whole-body severe jaundice, hepatosplenomegaly, kidney, and lymph node hemorrhages. The Colpodella sp.-specific 18S rRNA gene was detected using nested PCR. Interestingly, the DNA isolated from the blood of the tiger was found to be 100% similar to that of the tick by NCBI BLAST analysis. However, the DNA fragments isolated from the tiger's blood were 90.1% similar to the Colpodella sp. strain human erythrocyte parasite (HEP, MH208621) and 90.4% similar to the Colpodella sp. strain Heilongjiang (HLJ, KT364261). To investigate the species of ticks and ticks-carried Colpodella parasites in this region, the species of ticks obtained from the grasses outside the tiger enclosure and the species of Colpodella carried by ticks were identified. The DNA from ticks as well as that from the tick-borne Colpodella sp. were amplified from each tick using PCR followed by amplicon sequencing. In total 402 adult ticks samples were collected, among which 22 were positive for Colpodella sp. (5.5%), and the species were further determined by morphology, DNA sequencing and phylogenetic analyses. Interestingly, one Colpodella sp. was found to have 94.2% sequence similarities to the Colpodella sp. strain HEP (MH208621). This strain was previously reported to infect a woman in Yunnan, China. In addition, three Colpodella sp. showed 87-91% sequence similarities to the Colpodella sp. strain HLJ (KT364261), which was previously reported to infect human in Heilongjiang, China. This study disclosed the possibility of zoonotic transmission of Colpodella sp. by ticks in China. Finally, it provides a basis for urgently determining and monitoring the repertoire of ticks-borne piroplasmid pathogens, with the ultimate aim of strategic control.

Investigation of urine metabolome of BALB/c mouse infected with an avirulent strain of Toxoplasma gondii.

BACKGROUND: The protozoan parasite Toxoplasma gondii is a major concern for human and animal health. Although the metabolic understanding of toxoplasmosis has increased in recent years, the analysis of metabolic alterations through noninvasive methodologies in biofluids remains limited. METHODS: Here, we applied liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics and multivariate statistical analysis to analyze BALB/c mouse urine collected from acutely infected, chronically infected and control subjects. RESULTS: In total, we identified 2065 and 1409 metabolites in the positive electrospray ionization (ESI +) mode and ESI - mode, respectively. Metabolomic patterns generated from principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) score plots clearly separated T. gondii-infected from uninfected urine samples. Metabolites with altered levels in urine from T. gondii-infected mice revealed changes in pathways related to amino acid metabolism, fatty acid metabolism, and nicotinate and nicotinamide metabolism. CONCLUSIONS: This is the first study to our knowledge on urine metabolic profiling of BALB/c mouse with T. gondii infection. The urine metabolome of infected mouse is distinctive and has value in the understanding of Toxoplasmosis pathogenesis and improvement of treatment.

Identification of novel biofilm genes in avian pathogenic Escherichia coli by Tn5 transposon mutant library.

Avian pathogenic Escherichia coli (APEC) is the main pathogens that inflict the poultry industry. Biofilm as the pathogenic factors of APEC, which can enhance the anti-host immune system of APEC and improve its survival in the environment. In order to screen for new genes related to APEC biofilm. The APEC strain APEC81 was used to construct a mutant library by Tn5 insertion mutagenesis. Moreover the 28 mutant strains with severely weakened biofilm were successfully screened from 1500 mutant strains by crystal violet staining, in which 17 genes were obtained by high-efficiency thermal asymmetric interlaced PCR. The reported genes include 3 flagella genes (fliS, fliD, and fliR), 4 curli fimbriae genes (csgD, csgA, csgF, and csgG) and 3 type 1 fimbriae genes (fimA, fimD, and fimC). The novel genes include 3 coenzyme genes (gltA, bglX, and mltF) and 4 putative protein genes (yehE, 07045, 11735, 11255). To investigate whether these 17 genes co-regulate the biofilm, the 17 identified genes were deleted from APEC strain APEC81. The results showed that except for the 11735 and 11255 genes, the deletion of 15 genes significantly reduced the biofilm formation ability of APEC81 (P < 0.05). The result of rdar (red, dry and rough) colony morphology showed that curli fimbriae genes (csgD, csgA, csgF, and csgG) and other functional genes (fimC, glxK, yehE, 07045, and 11255) affected the colony morphology. In particular, the hypothetical protein YehE had the greatest influence on the biofilm. It was predicted to have the same structure as the type 1 fimbria protein. When yehE was deleted, the fimE transcription was up-regulated, and the fimA and fimB transcription were down-regulated, resulting in a decrease in type 1 fimbriae. Hence, the yehE mutant significantly reduced the biofilm and the adhesion and invasion ability to cells (P < 0.05). This study identified 5 novel genes (gltA, bglX, mltF, yehE, and 07045) related to biofilm formation and confirmed that yehE affects biofilm formation by type 1 fimbriae, which will benefit further study of the mechanism of biofilm regulation in APEC.

Taurine inhibits necroptosis helps to alleviate inflammatory and injury induced by Klebsiella infection.

Klebsiella infection is widely acknowledged to inflict severe inflammatory damage in bovines. Herein, we demonstrate significant death of EpH4-Ev cells incubated with Klebsiella. And compelling evidence shows that Klebsiella infection increases interactions between the Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and RIPK3, which promotes phosphorylation of RIPK3 and MLKL to induce necroptosis. However, these changes can be partially reversed by taurine and Nec-1s. Moreover, using taurine and Nec-1s to partially inhibit necroptosis significantly reduce TNF-α, IL-1ß and IL-6 levels and NAGase activity induced by Klebsiella infection. Taken together, taurine partially inhibits necroptosis induced by Klebsiella infection and hence alleviates inflammatory and injury in EpH4-Ev cells.

The Effects of the Solution-Focused Model on Anxiety and Postpartum Depression in Nulliparous Pregnant Women.

Background: Solution-focused model (SFM) is an intervention method that fully mobilizes patients' initiative through their potential. We aimed to investigate the effects of SFM on anxiety and postpartum depression (PPD) in nulliparous pregnant women compared with routine care services. Methods: We chose the mothers diagnosed as depressed or with depressive tendency by Edinburgh Postpartum Depression Scale (EPDS) at 28 weeks of gestation and divided them into the intervention and control groups. The control group only took the routine pregnancy healthy nursing, while the SFM group took the regular nursing and SFM counselling. Different assessments were conducted at 28 weeks of gestation, post-delivery, and post-intervention to evaluate the anxiety and depression levels of the patients. Finally, nursing satisfaction was evaluated by the nursing satisfaction questionnaire. Results: Compared with the control group, SFM could decrease the scores of anxiety and depression more effectively and influence sleep quality more positively. We also found that SFM resulted in significantly higher nursing satisfaction than that in the control group (p = 0.0046). Conclusion: In conclusion, SFM could effectively alleviate anxiety and PPD in nulliparous pregnant women.

Effect of nutritional and environmental conditions on biofilm formation of avian pathogenic Escherichia coli.

AIMS: To study the effects of environmental stress and nutrient conditions on biofilm formation of avian pathogenic Escherichia coli (APEC). METHODS AND RESULTS: The APEC strain DE17 was used to study biofilm formation under various conditions of environmental stress (including different temperatures, pH, metal ions, and antibiotics) and nutrient conditions (Luria-Bertani [LB] and M9 media, with the addition of different carbohydrates, if necessary). The DE17 biofilm formation ability was strongest at 25°C in LB medium. Compared to incubation at 37°C, three biofilm-related genes (csgD, dgcC, and pfs) were significantly upregulated and two genes (flhC and flhD) were downregulated at 25°C, which resulted in decreased motility. However, biofilm formation was strongest in M9 medium supplemented with glucose at 37°C, and the number of live bacteria was the highest as determined by confocal laser scanning microscopy. The bacteria in the biofilm were surrounded by a thick extracellular matrix, and honeycomb-like or rough surfaces were observed by scanning electron microscopy. Moreover, biofilm formation of the DE17 strain was remarkably inhibited under acidic conditions, whereas neutral and alkaline conditions were more suitable for biofilm formation. Biofilm formation was also inhibited at specific concentrations of cations (Na+ , K+ , Ca2+ , and Mg2+ ) and antibiotics (ampicillin, chloramphenicol, kanamycin, and spectinomycin). The real-time quantitative reverse transcription PCR showed that the transcription levels of biofilm-related genes change under different environmental conditions. CONCLUSIONS: Nutritional and environmental factors played an important role in DE17 biofilm development. The transcription levels of biofilm-related genes changed under different environmental and nutrient conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings suggest that nutritional and environmental factors play an important role in APEC biofilm development. Depending on the different conditions involved in this study, it can serve as a guide to treating biofilm-related infections and to eliminating biofilms from the environment.

Differential metabolism-associated gene expression of duck pancreatic cells in response to two strains of duck hepatitis A virus type 1.

Several outbreaks of duck hepatitis A virus type 1 (DHAV-1), which were characterized by yellow coloration and hemorrhage in pancreatic tissues, have occurred in China. The causative agent is called pancreatitis-associated DHAV-1. The mechanisms involved in pancreatitis-associated DHAV-1 infection are still unclear. Transcriptome analysis of duck pancreas infected with classical-type DHAV-1 and pancreatitis-associated DHAV-1 was carried out. Deep sequencing with Illumina-Solexa resulted in a total of 53.9 Gb of clean data from the cDNA library of the pancreas, and a total of 29,597 unigenes with an average length of 993.43 bp were generated by de novo sequence assembly. The expression levels of D-3-phosphoglycerate dehydrogenase, phosphoserine aminotransferase, and phosphoserine phosphatase, which are involved in glycine, serine, and threonine metabolism pathways, were significantly downregulated in ducks infected with pancreatitis-associated DHAV-1 compared with those infected with classical-type DHAV-1. These findings provide information regarding differences in expression levels of metabolism-associated genes between ducks infected with pancreatitis-associated DHAV-1 and those infected with classical-type DHAV-1, indicating that intensive metabolism disorders may contribute to the different phenotypes of DHAV-1-infection.

3'UTR SL-IV and DB1 Regions Contribute to Japanese Encephalitis Virus Replication and Pathogenicity.

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus that causes fatal neurological disease in humans, is one of the most important emerging pathogens of public health significance. JEV is maintained in an enzootic cycle and causes reproductive failure in pigs. Notably, the shift in JEV genotypes is not fully protected by existing vaccines, so the development of a candidate vaccine is urgently needed. In this study, we compared pathogenicity between Japanese encephalitis virus SA14 and BJB (isolated from humans in the 1970s) strains. We found that the BJB strain was attenuated in mice and that there was no case fatality rate. The growth rate of BJB was higher than SA14 virus in BHK-21 cells. Based on the sequence alignment of the viral genome between the SA14 and BJB virus strains, some mutations at sites 248, 254, 258, and 307 were observed in the 3' untranslated region (3'UTR). The 3'UTR of JEV plays a very important role in the viral life cycle. Furthermore, using a reverse genetic system, we conducted and rescued the parental JEV strain SA14 (T248, A254, and A258) and the mutant virus rSA14-3'UTRmut (T248C, A254G, A258G, and 307G). Through an analysis of the RNA secondary structure model of the 3'UTR, we discovered that the mutations of T248C, A254G, and A258G reduced the apiculus ring and increased the lateral ring significantly in the stem-loop structures IV (SL-IV) structure region of 3'UTR. Moreover, the insertion of 307G added a ring to the dumbbell structure 1 (DB1) structure region. Strikingly, these RNA secondary structure changes in 3'UTR of rSA14-3'UTRmut increased viral negative chain RNA production and enhanced the replication ability of the virus in BHK-21 cells. However, in vivo mouse experiments illustrated that the rSA14-3'UTRmut virus significantly decreased the neurovirulence of JEV. These results affirmed that the JEV SL-IV and DB1 regions play an important role in viral proliferation and pathogenicity. Taken together, we complement the study of RNA element function in the 3'UTR region of JEV by providing a new target for the rational design of live attenuated candidate vaccines and the increase of virus production.

Knowledge and attitude on donation of breast milk in hospitalized mothers / Conocimiento y actitud sobre la donación de leche materna en madres hospitalizadas

Objective: To understand the knowledge and attitude of breast milk donation among hospitalized mothers, and provide data to support the establishment and development of breast milk bank in China. Methods: By convenient sampling method, 200 hospitalized mothers were interviewed by ‘The Questionnaire on Knowledge and Attitude toward Breast Milk Donation in Hospitalized Mothers’ in Shanghai Sixth People's Hospital from September 2015 to November 2018. Results: The overall correct answer rate about the knowledge on breast milk donation was not high (29.23%). The average score of breast milk donation attitude was relatively low, with a score of 32.97 ± 4.30. Conclusion: The average correct answer rate for breast milk donation knowledge was relatively low, and the attitude on breast milk donation was not positive. The scope and intensity of breast milk donation are suggested to be expanded to promote the construction of breast milk banks. (AU)

Objetivo: Investigar el conocimiento y la actitud sobre la donación de leche materna entre las madres hospitalizadas, y aportar datos para respaldar el establecimiento y desarrollo del banco de leche materna en China. Métodos: Mediante el método de muestreo conveniente, se entrevistó a 200 madres hospitalizadas utilizando el Cuestionario de conocimiento y actitud sobre la donación de leche materna en madres hospitalizadas en el Hospital Popular Sixth de Shanghai desde septiembre de 2015 hasta noviembre de 2018. Resultados: Lasa de respuesta correcta global acerca del conocimiento de la donación de leche materna no fue alta (29,23%). La puntuación media de la actitud sobre la donación de leche materna fue relativamente baja: 32,97 ± 4,30. Conclusión: La tasa de respuesta correcta media para el conocimiento sobre la donación de leche materna fue relativamente baja, y la actitud sobre la donación de leche materna no fue positiva. El alcance y la intensidad de la donación de leche materna deben expandirse, para promover la construcción de bancos de leche materna. (AU)

The decreased exclusion of nuclear eccDNA: From molecular and subcellular levels to human aging and age-related diseases.

Extrachromosomal circular DNA (eccDNA) accumulates within the nucleus of eukaryotic cells during physiological aging and in age-related diseases (ARDs) and the accumulation could be caused by the declined exclusion of nuclear eccDNA in these states. This review focuses on the formation of eccDNA and the roles of some main factors, such as nuclear pore complexes (NPCs), nucleoplasmic reticulum (NR), and nuclear actin, in eccDNA exclusion. eccDNAs are mostly formed from non-coding DNA during DNA damage repair. They move to NPCs along nuclear actin and are excluded out of the nucleus through functional NPCs in young and healthy cells. However, it has been demonstrated that defective NPCs, abnormal NPC components and nuclear actin rods are increased in aged cells, various cancers and certain other ARDs such as cardiovascular diseases, premature aging, neurodegenerative diseases and myopathies. Therefore, mainly resulting from the increase of dysfunctional NPCs, the exclusion of nuclear eccDNAs may be reduced and eccDNAs thus accumulate within the nucleus in aging and the aforementioned ARDs. In addition, the protective function of non-coding DNA in tumorigenesis is further discussed.

Detection and molecular characterisation of intestinal parasites in the South China tiger Panthera tigris amoyensis (Hilzheimer).

Parasitic infections of the South China tigers in the Meihua Mountains have not been explored previously. Faeces of 22 South China tigers from the China Tiger Park in the Meihua Mountains were examined. Eggs of ascaridoid nematodes and oocysts of coccidia were detected by Mini-FLOTAC assay. Morphological observation and molecular characterisation of the oocysts were carried out. The prevalence of Toxascaris leonina (von Linstow, 1902) was 18% (4/22), and the highest egg per gram (EPG) count in the faeces was 27,150. The prevalence of Cystoisospora sp. was 45% (1 0/22) and the highest oocysts per gram (OPG) in the faeces was 6,000. In addition, we found one ascaridoid nematode in the South China tiger's faeces and was molecularly and morphologically identified as T. leonina. The oocysts in the faeces were sporulated in vitro and identified as Cystoisospora sp. Amplification of full-length internal transcribed spacers (ITS) resulted in sequences 1,622 bp long. Using the sequences, Cystoisospora sp. of the South China tiger was closest to Isospora belli (Wenyon, 1923) and Cystoisospora suis (Biester, 1934).