[The network investigation on knowledge, attitude and practice about COVID-19 of the residents in Anhui Province].

Objective: To analyze the current situation of the knowledge, attitudes and practice about COVID-19 of the residents in Anhui Province. Methods: Anonymous network sampling survey was carried out with an electronic questionnaire that designed by the questionnaire star, and a total of 4 016 subjects from Anhui province were investigated. The content of the survey includes that the basic information of subjects,the residents' knowledge, attitudes and practice about COVID-19, as well as their satisfaction with the prevention and control measures adopted by the government and health authorities and the suggestions on future prevention. The questionnaire doesn't involve any privacy information, and all questions were mandatory to ensure the response rate. Results: The M (P(25), P(75)) age the 4 016 subjects was 21 (19, 24) years old, and the ranging from 7 to 80 years old. The number of males was 1 431 (35.6%). Social networking tools such as WeChat and QQ were the main sources of epidemic information for residents (97.8%, 3 929 respondents). Residents had higher awareness rate of cough (99.5%,n=3 997) and fever (96.0%, n=3 857) symptoms, the transmission by droplets (99.5%, n=3 995), aerosol transmission (81.1%, n=3 258), and contact transmission (92.3%, n=3 708), but lower awareness of symptoms os muscle pain or fatigue (62.7%, n=2 518). 92.6% of the subjects (n=3 720) think that the outbreak was scary. In terms of psychological behavior scores, the results showed that female (9.38±4.81), the urban (9.37±5.02) and the medical workers (10.79±5.19) had a poorer mental health than the male (8.45±5.00), the rural (8.71±4.75) and the non-medical workers (the students: 8.85±4.83; public institude workers: 9.02±5.08; others: 8.97±5.39) (P<0.05). 71.9% of the residents (n=2 887) were satisfied with the local epidemic control measures. The residents took various of the measures to prevent and control the epidemic. The ratio of residents that could achieve "no gathering and less going out" , "wear masks when going out " and " do not go to crowded and closed places " was up to 97.4% (n=3 913), 93.6% (n=3 758) and 91.5% (n=3 673) respectively. Conclusion: The residents in Anhui province have a good KAP about COVID-19, yet it is necessary to strengthen the community publicity, the mental health maintenance of residents and students' health education.

[The network investigation on knowledge, attitude and practice about Novel coronavirus pneumonia of the residents in Anhui Province].

Objective: To analyze the current situation of the knowledge, attitudes and practice about Novelcoronavirus pneumonia (NCP) of the residents in Anhui Province. Methods: Anonymous network sampling survey was carried out with an electronic questionnaire that designed by the questionnaire star, and a total of 4016 subjects from Anhui province were investigated. The content of the survey includes that the basic information of subjects,the residents' knowledge, attitudes and practice about NCP, as well as their satisfaction with the prevention and control measures adopted by the government and health authorities and the suggestions on future prevention. The questionnaire doesn't involve any privacy information, and all questions were mandatory to ensure the response rate. Results: The M (P(25), P(75)) age the 4016 subjects was 21 (19, 24), and the ranging from 7 to 80 years old. The number of males was1431(35.6%). Social networking tools such as WeChat and QQ were the main sources of epidemic information for residents (97.8%, 3 929 respondents). Residents have a high awareness rate of the main symptoms, transmission routes, using of masks, hand washing and treatment information of NCP, while a low awareness rate of the atypical symptoms. 92.6% of the subjects (n=3 720) think that the outbreak was scary. In terms of psychological behavior scores, the results showed that female (9.38±4.81), the urban (9.37±5.02) and the medical workers (10.79±5.19) had a poorer mental health than the male (8.45±5.00) , the rural (8.71±4.75) and the non-medical workers (the students: 8.85±4.83; public institude workers: 9.02±5.08; others: 8.97±5.39) (P < 0.05). 71.9% of the residents (n=2 887)were satisfied with the local epidemic control measures. The residents took various of the measures to prevent and control the epidemic. The ratio of residents that could achieve "no gathering and less going out" , "wear masks when going out" and "do not go to crowded and closed places" was up to 97.4% (n=3 913), 93.6% (n=3758) and 91.5% (n=3 673) respectively. Conclusion: The residents in Anhui province have a good KAP about NCP, yet it is necessary to strengthen the community publicity, the mental health maintenance of residents and students' health education.

Molecular cloning, functional characterization and phylogenetic analysis of TRAIL in Japanese pufferfish Takifugu rubripes.

In this study, the complementary DNA (cDNA) of Japanese pufferfish Takifugu rubripes tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) was cloned by reverse-transcription PCR. The open reading frame of the TRAIL consisted of 870 bases. The deduced amino-acid sequence of the TRAIL showed a high homology with the sequences of other teleosts. Recombinant soluble TRAIL was fused with a small ubiquitin-related modifier gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). In vitro, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide (MTT) assay indicated that the purified soluble TRAIL was able to induce apoptosis of Jurkat and HeLa cells in a dose-dependent manner.

Analysis of transcription factors regulating induction of indoleamine 2,3-dioxygenase by IFN-gamma.

IFN-gamma treatment of the human carcinoma cell line ME180 causes cell death due to induction of indoleamine 2,3-dioxygenase (IDO) and resulting starvation for tryptophan. A mutant cell line 3B6A derived from ME180 was resistant to IFN-gamma because of loss of IDO activity. Cotransfecting an IDO promoter-chloramphenicol acetyl transferase (CAT) construct with IFN regulatory factor-1 (IRF-1) resulted in induction of CAT activity in both ME180 and 3B6A cells even in the absence of IFN-gamma. This induction was reduced by cotransfection with IRF-2. However, IRF-1 was not able to restore IDO activity, suggesting a possible repressor site outside the IDO promoter region. Stat1alpha (p91) restored both CAT and IDO activities in 3B6A cells following IFN-gamma treatment. 3B6A cells doubly treated with IFN-gamma and IFN-alpha or IFN-beta restored IDO activity, although neither cytokine on its own could induce IDO. Western blot analysis showed that both constitutive expression and induction of Stat1alpha by IFN-gamma were reduced in 3B6A cells, and double treatment of IFN-gamma with IFN-alpha or IFN-beta restored the expression level of Statla. Electrophoretic mobility shift assays indicated that Stat1 binds to the IFN-gamma-activated sequence (GAS) region in the IDO promoter in ME180 cells following IFN-gamma treatment. Our results indicated that the defect in 3B6A cells was reduced expression of Stat1alpha and that IRF-1, NF-kappaB, and PKR were all involved to some extent in the induction of IDO following IFN-gamma treatment.

Antitumor activity of recombinant adenoviral vectors expressing murine IFN-alpha in mice injected with metastatic IFN-resistant tumor cells.

Recent studies have shown that gene therapy with type I interferon (IFN) in an adenovirus vector is a powerful tool to suppress the growth of human tumors transplanted in immune-deficient mice. However, in these studies the host immune-mediated effects, which may be important in mediating the long-term control of tumor growth by these cytokines, was not studied. In this paper, we evaluate the antitumor efficacy of different adenoviral vectors containing mouse IFN-alpha genes (i.e., a first-generation replication-defective vector containing IFN-alpha1 and two different second-generation vectors containing IFN-alpha2) in immunocompetent DBA/2 mice transplanted with highly metastatic Friend leukemic cells resistant in vitro to type I IFN. We found that injection of all the different adenovirus vectors containing mouse IFN-alpha( genes resulted in a marked antitumor response in mice transplanted either subcutaneously or intravenously with IFN-resistant Friend leukemic cells compared to tumor-bearing animals inoculated with a control vector. Tumor growth inhibition after injection of IFN-adenovirus vectors was associated with a prolonged presence of high IFN levels in the sera of the injected mice. Suppression of metastatic tumor growth was also observed after a single injection of the IFN--adenovirus recombinant vectors, whereas a comparable antitumor response generally required several injections of high doses of IFN. Altogether, these results demonstrate that IFN--adenoviral vectors can efficiently inhibit metastatic tumor growth by host-mediated mechanisms and suggest that adenovirus-mediated IFN-alpha gene therapy may represent an attractive alternative to the conventional clinical use of this cytokine, which generally requires multiple injections of high IFN doses for a prolonged period of time.

Suppression of Frizzled-2-mediated Wnt/Ca²âº signaling significantly attenuates intracellular calcium accumulation in vitro and in a rat model of traumatic brain injury.

Traumatic brain injury (TBI) can dramatically increase levels of intracellular calcium (Ca²âº). The association between Wnt5a/Frizzled-2 (wingless-type mouse mammary tumor virus integration site family member 5a/Fzd2) signaling and Ca²âº cellular homeostasis in lower vertebrates has been well documented. However, little is known about Wnt5a/Fzd2 signaling in mammalian nerve cells, or whether Ca²âº accumulation after TBI is mediated through this pathway. We hypothesized that an activated Wnt5a/Fzd2 pathway following TBI may play a role in Ca²âº overloading. To elucidate the influence of Fzd2 and the Wnt5a signal transduction pathway on an increase in intracellular Ca²âº, we assessed the expression of Wnt5a/Fzd2 in rat hippocampal cells both in vitro and in vivo. We found that transfection of the rat Fzd2 gene in rat neonatal hippocampal astrocytes significantly increased gene expressions of both Wnt5a and Fzd2 by fourfold when compared to non-transfected cells (P<0.01 in both cases). Expressions of the proteins Wnt5a and Fzd2 were significantly increased approximately two- and threefold, respectively, when compared to non-transfected control cells (P<0.01 in both cases). Moreover, intracellular Ca²âº, as manifested by the fluorescent intensity of the intracellular Ca²âº indicator Fluo-3/AM, was significantly increased by 1.75-fold (P<0.01). The blocking of Fzd2 signaling using Stealth RNAi markedly inhibited the elevated gene and protein expression of Wnt5a in the transfected cells by two- and fourfold, respectively (P<0.01), and suppressed intracellular Ca²âº by 1.5-fold (P<0.01). Furthermore, in vivo, we demonstrated that TBI-induced dramatic upregulation of gene and protein expression of Wnt5a/Fzd2 by two- and fivefold (P<0.01) in injured hippocampi, and intracellular Ca²âº increased in isolated injured hippocampal cells. Whereas, the in vivo blocking of Fzd2 signaling by hippocampal delivery of Stealth RNAi and Invivofectamine significantly suppressed the increased gene and protein expression of Wnt5a and Fzd2 induced by TBI by 1- to 3.5-fold (P<0.01) and also inhibited Ca²âº accumulation by 1.5-fold (P<0.01). These findings demonstrated that the Wnt5a/Fzd2 signaling pathway contributed to increasing intracellular Ca²âº in nerve cells under physiological and pathological conditions. Furthermore, our findings provide evidence that specifically expressed components of this signal pathway, such as Wnt5a and Fzd2, are potential therapeutic targets following brain trauma.

Indoleamine 2,3-dioxygenase production by human dendritic cells results in the inhibition of T cell proliferation.

Dendritic cells (DCs) play a key role in the activation and regulation of B and T lymphocytes. Production of indoleamine 2, 3-dioxygenase (IDO) by macrophages has recently been described to result in inhibition of T cell proliferation through tryptophan degradation. Since DCs can be derived from monocytes, we sought to determine whether DCs could produce IDO which could potentially regulate T cell proliferation. Northern blot analysis of RNA from cultured monocyte-derived human DC revealed that IDO mRNA was induced upon activation with CD40 ligand and IFN-gamma. IDO produced from activated DCs was functionally active and capable of metabolizing tryptophan to kynurenine. Activated T cells were also capable of inducing IDO production by DCs, which was inhibited by a neutralizing Ab against IFN-gamma. DC production of IDO resulted in inhibition of T cell proliferation, which could be prevented using the IDO inhibitor 1-methyl-dl -tryptophan. These results suggest that activation of DCs induces the production of functional IDO, which causes depletion of tryptophan and subsequent inhibition of T cell proliferation. This may represent a potential mechanism for DCs to regulate the immune response.