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Background & aims Although the prevalence of nonalcoholic fatty liver disease (NAFLD) has risen dramatically to 25% of the adult population worldwide, there are as yet no approved pharmacological interventions for the disease due to uncertainty about the underlying molecular mechanisms. It is known that mitochondrial dysfunction is an important factor in the development of NAFLD. Mitochondrial antiviral signaling protein (MAVS) is a critical signaling adaptor for host defenses against viral infection. However, the role of MAVS in mitochondrial metabolism during NAFLD progression remains largely unknown. Approach & results Based on expression analysis, we identified a marked downregulation of MAVS in hepatocytes during NAFLD progression. By employing MAVS global knockout and hepatocyte-specific MAVS knockout mice, we found that MAVS is protective against diet-induced NAFLD. MAVS deficiency induces extensive mitochondrial dysfunction during NAFLD pathogenesis which was confirmed as impaired mitochondrial respiratory capacity and membrane potential. Metabolomics data also showed the extensive metabolic disorders after MAVS deletion. Mechanistically, MAVS interacts with the N-terminal stretch of voltage-dependent anion channel 2 (VDAC2), which is required for the ability of MAVS to influence mitochondrial function and hepatic steatosis. Conclusions In hepatocytes, MAVS plays an important role in protecting against NAFLD by helping to regulate healthy mitochondrial function. These findings provide new insights regarding the metabolic importance of conventional immune regulators and support the possibility that targeting MAVS may represent a new avenue for treating NAFLD.

Multiple SARS-CoV-2 variants of concern (VOCs) have been emerging and some have been linked to an increase in case numbers globally. However, there is yet a lack of understanding of the molecular basis for the interactions between the human ACE2 (hACE2) receptor and these VOCs. Here we examined several VOCs including Alpha, Beta, and Gamma, and demonstrate that five variants receptor-binding domain (RBD) increased binding affinity for hACE2, and four variants pseudoviruses increased entry into susceptible cells. Crystal structures of hACE2-RBD complexes help identify the key residues facilitating changes in hACE2 binding affinity. Additionally, soluble hACE2 protein efficiently prevent most of the variants pseudoviruses. Our findings provide important molecular information and may help the development of novel therapeutic and prophylactic agents targeting these emerging mutants., The SARS-CoV-2 spike (S) protein mediates viral entry by binding of its receptor-binding domain (RBD) to the human angiotensin-converting enzyme 2 (ACE2) receptor and mutations of the S protein may have a great impact on virus transmissibility. Here, the authors characterize the interactions of six different SARS-CoV-2 RBD variants among them Alpha, Beta and Gamma and present crystal structures of these ACE2-RBD complexes.

Background and aims NAFLD is the most prevalent chronic liver disease worldwide, but no effective pharmacological therapeutics are available for clinical use. NASH is the more severe stage of NAFLD. During this progress, dysregulation of endoplasmic reticulum (ER)-related pathways and proteins is one of the predominant hallmarks. We aimed to reveal the role of ring finger protein 5 (RNF5), an ER-localized E3 ubiquitin-protein ligase, in NASH and to explore its underlying mechanism. Approach and results We first inspected the expression level of RNF5 and found that it was markedly decreased in livers with NASH in multiple species including humans. We then introduced adenoviruses for Rnf5 overexpression or knockdown into primary mouse hepatocytes and found that palmitic acid/oleic acid (PAOA)-induced lipid accumulation and inflammation in hepatocytes were markedly attenuated by Rnf5 overexpression but exacerbated by Rnf5 gene silencing. Hepatocyte-specific Rnf5 knockout significantly exacerbated hepatic steatosis, inflammatory response, and fibrosis in mice challenged with diet-induced NASH. Mechanistically, we identified 3-hydroxy-3-methylglutaryl CoA reductase degradation protein 1 (HRD1) as a binding partner of RNF5 by systematic interactomics analysis. RNF5 directly bound to HRD1 and promoted its lysine 48 (K48)-linked and K33-linked ubiquitination and subsequent proteasomal degradation. Furthermore, Hrd1 overexpression significantly exacerbated PAOA-induced lipid accumulation and inflammation, and short hairpin RNA-mediated Hrd1 knockdown exerted the opposite effects. Notably, Hrd1 knockdown significantly diminished PAOA-induced lipid deposition, and up-regulation of related genes resulted from Rnf5 ablation in hepatocytes. Conclusions These data indicate that RNF5 inhibits NASH progression by targeting HRD1 in the ubiquitin-mediated proteasomal pathway. Targeting the RNF5-HRD1 axis may provide insights into the pathogenesis of NASH and pave the way for developing strategies for NASH prevention and treatment.

BACKGROUND: Transarterial bland embolization (TABE) is widely used to treat the spontaneous rupture of hepatocellular carcinoma (HCC), and can lead to ischemic necrosis of the tumor. In this study, we used the propensity-score matching (PSM) method to compare the initial responses of treatment-naïve HCC patients to TABE and drug-eluting beads-transarterial chemoembolization (DEB-TACE), and the safety of these treatments. METHODS: Patients with treatment-naïve HCC, who had been admitted to 2 medical centers from January 2016 to December 2020, were enrolled as the research subjects. The data of 26 patients treated with TABE for ruptured HCC and 52 patients treated with DEB-TACE for primary HCC were collected according to our inclusion and exclusion criteria, and a PSM analysis was conducted to assess the safety and effectiveness of these two interventional techniques 1 month postoperatively. RESULTS: In relation to ruptured HCC, TABE had a hemostatic success rate of 97.0%. Before PSM, the TABE group had a larger maximum tumor diameter (P0.05). The multivariate regression analysis showed that tumor size was an independent predictor of the objective response rate (ORR) [odds ratio (OR): 3.312; 95% CI: 0.152–5.944; P0.05). The incidences of post-embolization syndrome (PES) and 48-h hepatotoxicity were significantly lower in the TABE group than the DEB-TACE group (both P0.05). CONCLUSIONS: TABE is highly effective at managing hemorrhage from ruptured HCC. The initial therapeutic response of HCC to TABE is similar to that to DEB-TACE; however, TABE is associated with lower hepatotoxicity and fewer adverse effects, which paves the way for subsequent treatments and systemic therapies.

Sepsis caused acute lung injury (ALI) is a kind of serious disease in critically ill patients with very high morbidity and mortality. Recently, it has been demonstrated that Golgi is involved in the process of oxidative stress. However, whether Golgi stress is associated with oxidative stress in septic induced acute lung injury has not been elucidated. In this research, we found that lipopolysaccharide (LPS) induced oxidative stress, apoptosis, inflammation and Golgi morphology changes in acute lung injury both in vivo and in vitro. The knockout of heme oxygenase-1(HO-1) aggravated oxidative stress, inflammation, apoptosis and reduced the expression of Golgi matrix protein 130 (GM130), mannosidase Ⅱ, Golgi-associated protein golgin A1 (Golgin 97), and increased the expression of Golgi phosphoprotein 3 (GOLPH3), which caused the fragmentation of Golgi. Furtherly, the activation of hypoxia inducible factor-1α (HIF-1α)/HO-1 pathway, attenuates Golgi stress and oxidative stress by increasing the levels of GM130, mannosidase Ⅱ, Golgin 97, and decreasing the expression of GOLPH3 both in vivo and in vitro. Therefore, the activation of HO-1 plays a crucial role in alleviating sepsis-induced acute lung injury by regulating Golgi stress, oxidative stress, which may provide a therapeutic target for the treatment of acute lung injury., Graphical abstract Image 1

The virus caused coronavirus disease 2019 (COVID-19 virus), also called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is the seventh coronavirus that can infect humans, and belongs to the genus β-coronavirus in the family Coronaviridae . As of 5 November 2020, SARS-CoV-2 has caused >47000000 confirmed cases and >1200000 related deaths in 219 countries and regions, bringing great challenges to global public health. Currently, there is no approved therapeutics or vaccines for the treatment of this disease. Several studies suggested SARS-CoV-2 might have originated from bats based on phylogenetic analysis, but the intermediate host of the virus is still unknown. Besides humans, cats, dogs, tigers, lions, minks, and other species have been reported to be infected by SARS-CoV-2. Several studies have reported pangolins as the only other mammalian species carrying coronaviruses related to SARS-CoV-2 besides bats, and suggested pangolins might be the intermediate host of SARS-CoV-2. Previously, we and other groups identified the angiotensin converting enzyme 2 (ACE2), the receptor of SARS-CoV, also functions as the entry receptor of SARS-CoV-2 and is recognized by the receptor binding domain (RBD) of the spike protein (S). Then, we elucidated the interaction between SARS-CoV-2 RBD and human ACE2 (hACE2) or cat ACE2 (cACE2), and also found that SARS-CoV-2 had broad potential host range, including domestic animals, companion pets and wild animals. In this study, we investigated the binding features of SARS-CoV-2 and two pangolin coronaviruses (pangolin-CoVs, GX/P2V/2017 and GD/1/2019) that recognize the receptors of both pangolin ACE2 (pACE2) and hACE2 using surface plasmon resonance (SPR) and structural methods. We further determined the crystal structure of SARS-CoV-2 S (RBD) in complex with pACE2 at a 2.3 A resolution, revealing the similarity in the binding mode between SARS-CoV-2 RBD to hACE2 and to cACE2. Interestingly, we found that the SARS-CoV-2 RBD-pACE2 complex is more similar to the SARS-CoV-2 RBD-hACE2 complex than to SARS-CoV-2 RBD-cACE2 complex. Furthermore, we modeled the interactions of GX/P2V/2017 RBD and GD/1/2019 RBD bound to pACE2 and hACE2, respectively, and found that both viruses adopted similar binding mode as SARS-CoV-2 RBD to pACE2 and hACE2. However, crystal structure and homology modeling implied that the interaction between the three RBDs and pACE2 were slightly weaker than their respective binding to hACE2, which was consistent with the receptor-RBD protein interaction measured by SPR. These results could facilitate better understanding of SARS-CoV-2 evolution, indicate the potential of pangolin-CoVs to enter human population that may lead to another outbreak, and highlight the importance of monitoring pangolin CoVs to prevent possible spillovers.

BACKGROUND: Thoracoscopic resection of small pulmonary nodules (SPNs) is challenging. Accurate preoperative computed tomography-guided localization of SPNs is key to successful rection. The aim of the present study was to evaluate the clinical value of a novel localization needle and methylene blue staining combined with surgical glue (MBSG) and to explore the risk factors for post-localization complications. METHODS: This prospective, non-randomized controlled study was conducted on 110 patients who received either MBSG or novel needle localization prior to video-assisted thoracoscopic surgery (VATS) from January 2019 to December 2019 at Shenzhen People’s Hospital. The primary endpoints were the safety and the success rates of the 2 localization techniques. The secondary endpoints were operative time and feasibility. RESULTS: The 110 patients were categorized into 2 groups: the MBSG group (n=84) and the pulmonary nodule localization needle group (n=26). The success rate of pre-VATS localization was 100% in both groups. No deaths or serious complications occurred during localization. The rates of pneumothorax, pulmonary hemorrhage, and localization-induced cough were 38.1%, 25%, and 7.14%, respectively, in the MBSG group, and 26.92%, 19.23%, and 0%, respectively, in the pulmonary nodule localization needle group. Differences between the 2 groups were not statistically significant (P>0.05). Total complication rate and the incidence of pain were significantly lower in the pulmonary nodule localization needle group (χ(2)=4.441 and 4.295, respectively; P0.05). Dye diffusion occurred in 2 patients in the MBSG group; however, it had no impact on VATS or on the pathological analysis. Neither displacement nor dislocation was observed in the pulmonary nodule localization needle group. Logistic regression analysis showed that the localization technique was an independent risk factor for total complications (odds ratio: 2.634, 95% confidence interval: 1.022–6.789, P

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease with high mortality (12%–30%). The mechanism by which the SFTS bunyavirus (SFTSV) causes severe illness remains unclear. To evaluate the phenotypic and functional characteristics of the NK cell subsets in SFTS patients, twenty-nine SFTS patients were sequentially sampled from admission until recovery. Phenotypic and functional characteristics of NK cell subsets in circulating blood were analysed via flow cytometry. Then, correlations between NK cell subset frequencies and the SFTS index (SFTSI) were evaluated in all SFTS patients (15 mild, 14 severe) upon admission. The frequencies of CD56dimCD16+ NK cells were greatly decreased in early SFTSV infection and were negatively correlated with disease severity. Additionally, higher Ki-67 and granzyme B expression and relatively lower NKG2A expression in CD56dimCD16+ NK cells were observed in acute infection. Moreover, the effector function of CD56dim NK cells was increased in the acute phase compared with the recovery phase in nine severe SFTS patients. Additionally, interleukin (IL)-15, interferon (IFN)-α, IL-18 and IFN-γ secretion was markedly increased during early infection. Collectively, despite depletion of CD56dimCD16+ NK cells, activation and functional enhancement of CD56dimCD16+ NK cells were still observed, suggesting their involvement in defence against early SFTSV infection.

Ovarian cancer is the most lethal gynecological tumor, and the 5-year survival rate is only ~40%. The poor survival rate is due to cancer diagnosis at an advanced stage, when the tumor has metastasized. A better understanding of the molecular pathogenesis of tumor growth and metastasis is needed to improve patient prognosis. MicroRNAs (miRs) regulate carcinogenesis and development of cancers. However, the role of miR-508-3p in ovarian cancer remains largely unknown. Thus, the present study aimed to investigate the possible functions of miR-508-3p in the modulation of development of ovarian cancer. The results of the present study demonstrated that miR-508-3p mimics inhibited ovarian cancer cell proliferation, migration and invasion. Reporter gene assay results demonstrated that miR-508-3p suppressed cancer cell proliferation by directly targeting the 3′-untranslated region (UTR) of cyclin A2 (CCNA2) and suppressed migration and invasion by directly targeting the 3′-UTR of matrix metallo-proteinase 7 (MMP7). In addition, high CCNA2 and MMP7 expression levels were associated with low miR-508-3p expression in ovarian cancer tissues. Furthermore, miR-508-3p and CCNA2 were independent predictors for overall survival in patients with ovarian cancer. To the best of our knowledge, this is the first study to demonstrated that miR-508-3p suppressed ovarian cancer development by directly targeting CCNA2 and MMP7. The results of this study suggested the potential value of miR-508-3p and CCNA2 as prognostic indicators and therapeutics for ovarian cancer.

Most of the currently developed Ga2O3-based solar blind photodetectors exhibit unexpected high persistent photoconductive gain at the expense of low response speed, and thus, the suppression of carrier trapping remains challenging. In this work, we demonstrated amorphous gallium oxynitride (GaON) based ultraviolet photodetectors with tunable spectral response and enhanced response speed by in-situ anion engineering with reactive sputtering technique. The tunable spectral response from 4.95 to 4.37 eV is a result of bandgap narrowing effect, attributed to the elevation of the valence band maximum (VBM) by the hybridization of N 2p and O 2p states and the enhanced p-d repulsion. The constructed GaON PDs with a proper nitrogen composition exhibit remarkably reduced dark current and a fast response time of about 100 µs. Oxygen vacancies are deactivated by the lift-up of VBM so that slow carrier detrapping processes are suppressed, resulting in the reduced persistent photoconductivity and improved response spe...

This study aimed to evaluate the safety and effectiveness of CT-guided percutaneous cryoablation for treatment of painful osteolytic bone metastases. A total of 26 patients (36 bone metastases) treated with CT-guided percutaneous cryoablation between May 2012 and June 2016 were enrolled in this retrospective study. All procedures were performed under local anesthesia. A visual analog scale (VAS) was used to evaluate pain before the procedure and at 1 day, 1 month, 3 months, and 6 months after the procedure. Complications during and after the procedure were recorded and graded by the Clavien-Dindo classification. The mean VAS pain score was 7.1 ± 1.1 (range, 4-10) before cryoablation. It was significantly lower at all timepoints after treatment: 2.1 ± 1.7 (P 0.0001) at 1 day after treatment, 1.3 ± 1.8 (P 0.0001) at 1 month, 1.6 ± 1.7 (P 0.0001) at 3 months, and 1.8 ± 1.3 (P 0.0001) at 6 months. The response rates were 91.7%, 94.4%, 91.7%, and 94.4%, respectively, at 1 day, 1 month, 3 months, and 6 months after cryoablation; the complete response rates were 22.2%, 41.7%, 36.1%, and 22.2%, respectively. Adverse events (skin frostbite, nerve injury, pathologic fracture) occurred in 3 patients. CT-guided percutaneous cryoablation under local anesthesia appears to be a safe and effective treatment for painful osteolytic bone metastases. Prospective clinical trials on large samples needed to confirm this conclusion.

Background: Kikuchi-Fujimoto disease (KFD), also named Histiocytic necrotizing lymphadenitis (HNL), is a rare and self-limited necrotizing lymphadenitis, most commonly affects young Asian female. KFD is characterized by lymphadenopathy, cytopenia, fever, rashes, and weight loss, however, hemophagocytic syndrome and pleural effusion are rarely reported. Case Presentation: We present the case of a 61-year-old man with clinical features including fever, lymphadenopathy, purpura, arthralgia, pleural effusion and pulmonary infection. He had cervical lymphnodebiopsy and the pathology is consistent with KFD, laboratory tests and clinical manifestations were consistent with hemophagocytic syndrome (HLH). After initial treatment including corticosteroid and anti-infection drugs, the patient showed improvement but soon deteriorated. Conclusion: KFD is a self-limited disease with spontaneous resolution. However, the cases of KFD with HLH may have a fatal course. Early appropriate intensive immunosuppressive therapy and strong anti-infection therapy could be beneficial for the treatment outcome.

Background/Aims Microvascular invasion (MVI) is an established risk factor for hepatocellular carcinoma (HCC). However, prediction models that specifically focus on the individual prognoses of HCC patients with MVI is lacking. Methods A total of 385 HCC patients with MVI were randomly assigned to training and validation cohorts in a 2:1 ratio. The outcomes were disease-free survival (DFS) and overall survival (OS). Prognostic nomograms were established based on the results of multivariate analyses. The concordance index (C-index), calibration plots and Kaplan-Meier curves were employed to evaluate the accuracy, calibration and discriminatory ability of the models. Results The independent risk factors for both DFS and OS included age, tumor size, tumor number, the presence of gross vascular invasion, and the presence of Glisson’s capsule invasion. The platelet-to-lymphocyte ratio was another risk factor for OS. On the basis of these predictors, two nomograms for DFS and OS were constructed. The C-index values of the nomograms for DFS and OS were 0.712 (95% confidence interval [CI], 0.679 to 0.745; p

Triclosan (TCS) was quantitatively analyzed for the first time in chicken samples using immunoaffinity cleanup followed by indirect competitive enzyme-linked immunosorbent assay (icELISA). Monoclonal antibodies (mAbs) against TCS were prepared. The mAb 6E1 showed high sensitivity and specificity in phosphate-buffered saline buffer, with a half maximal inhibition concentration value of 1.77 ng mL−1 and a limit of detection of 0.09 ng mL−1. The average recovery in spiked chicken muscle samples was 84.4–114.8%, with a relative standard deviation below 8.1%. To validate the developed icELISA method, samples spiked with different levels of TCS were analyzed by icELISA and ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). The consistency of results detected by icELISA and UPLC–MS/MS demonstrated that immunoaffinity cleanup followed by icELISA could also be used for other food matrices.

ODV-E66 is a major envelope proteins of baculovirus occlusion derived virus (ODV) with chondroitinase activity. Here, we studied the roles of ODV-E66 during Helicoverpa armigera nucleopolyhedrovirus (HearNPV) primary infection. ODV-E66 is a late viral protein dispensable for BV production and ODV morphogenesis. Deletion of odv-e66 had a profound effect on HearNPV oral infectivity in 4th instar larvae with a 50% lethal concentration (LC50) value of 26 fold higher than that of the repaired virus, compared to in 3rd instar larvae. Calcofluor white, an agent which destroys the peritrophic membrane (PM), could rescue the oral infectivity of odv-e66 deleted HearNPV, implying the PM may be the target of ODV-E66. In vitro assays showed HearNPV ODV-E66 has chondroitinase activity. Electron microscopy demonstrated that odv-e66 deletion alleviated the damage to the PM caused by HearNPV infection. These data suggest an important role of ODV-E66 in the penetration of the PM during oral infection.

A previous study reported that histone methyltransferase SETD3 is up-regulated in tumor tissues of hepatocellular carcinoma (HCC) and is associated with the growth of HCC. However, the clinical significance and the effect of SETD3 on HCC metastasis remain unclear. In the present study, both the protein and mRNA expression levels of SETD3 were measured in a larger cohort of HCC patients. The results showed that the protein level of SETD3 in HCC tissues was significantly higher than that in non-tumorous tissues, which was inconsistent with the mRNA expression level of SETD3. The high protein level of SETD3 in HCC tissues was significantly associated with male gender, poor pathological differentiation, liver cirrhosis and unfavorable prognosis of HCC patients. Subsequently, we demonstrated that SETD3 could be regulated at post-transcriptional step by a couple of miRNAs (miR-16, miR-195 and miR-497). Additionally, in vitro and in vivo experiments revealed that SETD3 played opposing roles in proliferation and metastasis of HCC: promoting proliferation but inhibiting metastasis. Mechanistic experiments revealed that doublecortin-like kinase 1 (DCLK1) was a downstream target of SETD3. SETD3 could increase the DNA methylation level of DCLK1 promoter to inhibit the transcription of DCLK1. Further study revealed that DCLK1/PI3K/matrix metalloproteinase (MMP) 2 (MMP-2) was an important pathway that mediated the effect of SETD3 on HCC metastasis. In conclusion, the present study revealed that SETD3 is associated with tumorigenesis and is a promising biomarker for predicting the prognosis of HCC patients after surgical resection. In addition, SETD3 plays inhibitory role in HCC metastasis partly through DCLK1/PI3K/MMP-2 pathway.

We report the isolation of Tamdy virus from Hyalomma asiaticum ticks in northwest China and serologic evidence of human Tamdy virus infection in the same region. These findings highlight the need to further investigate a potential causal relationship between Tamdy virus and febrile illnesses of unknown etiology in that region.

There is little information on pollen morphological analysis of papaya cultivars with different flesh color and origins. We observed pollen morphological characters of 17 papaya cultivars through scanning electron microscopy. The results showed that papaya pollen grains were monads, tricolporate, and small and medium-sized. The pollen grains were oblate-spheroidal to spheroidal. Foveolate, reticulate, and fossulate exine ornamentation were observed. Colpi width had maximum coefficient of variation of 25%; Shannon diversity index of all qualitative characters reached over 0.8. Using cluster analysis, 17 papaya cultivars were divided into three groups; there was no relationship between flesh colors and origins. The study findings suggest that pollen morphological analysis is valuable to provide references for breeding and genetic improvement of papaya.

SARS-CoV-2 can infect many domestic animals, including dogs. Herein, we show that dog angiotensin-converting enzyme 2 (dACE2) can bind to the SARS-CoV-2 spike (S) protein receptor binding domain (RBD), and that both pseudotyped and authentic SARS-CoV-2 can infect dACE2-expressing cells. We solved the crystal structure of RBD in complex with dACE2 and found that the total number of contact residues, contact atoms, hydrogen bonds and salt bridges at the binding interface in this complex are slightly fewer than those in the complex of the RBD and human ACE2 (hACE2). This result is consistent with the fact that the binding affinity of RBD to dACE2 is lower than that of hACE2. We further show that a few important mutations in the RBD binding interface play a pivotal role in the binding affinity of RBD to both dACE2 and hACE2. Our work reveals a molecular basis for cross-species transmission and potential animal spread of SARS-CoV-2, and provides new clues to block the potential transmission chains of this virus., Many domestic animals, among them dogs, have been infected with SARS-CoV-2 during the COVID-19 pandemic. Here, the authors present the crystal structure of the SARS-CoV-2 spike protein receptor binding domain (RBD) bound to its receptor, dog angiotensin-converting enzyme 2 (dACE2), and show that the RBD N501Y mutation increases the infectivity and host range of SARS-CoV-2, which highlights the need to monitor emerging SARS-CoV-2 mutations in domestic animals.

Background Canine distemper caused by canine distemper virus that belongs to the Morbillivirus genus of the Paramyxoviridae family is still a global epidemic significant infectious disease, especially in pet dogs in China and serious harm to the development of the dog industry. It has been known that apoptosis caused by the canine distemper virus can show in culture cells, lymphoid tissues, and the cerebellum. However, its occurrence in brain tissue cells remains unclear. To investigate the relationship among canine distemper infecting brain tissues, apoptosis in brain tissue cells, and demyelinating pathogenesis was investigated. Methods 16 naturally infected dogs that exhibited clinical signs of CD and tested positive for the anti-CDV monoclonal antibody and six healthy dogs that served as the control, were used in the research. Brain specimens were divided into the cerebrum, brain stem, and cerebellum embedded in paraffin and made the sections respectively. Approximately 5 µm-thick sections were stained by hematoxylin–eosin, methyl green pyronin, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling technique, and immunohistochemistry. CDV nucleocapsid protein was detected by immune streptavidin-biotinylated peroxidase complex. Results Alterations in the brain tissues of CDV-infected dogs involved both various cells and nerve fibers. CDV had varying degrees of cytotropism to all brain tissue cells; apoptosis also occurred in all brain cells, especially in the endothelia of cerebral vessels, astrocytes, oligodendrocytes, and ependymal cells, the more serious infection, the more obvious apoptosis. Serious infections also involved the pyramidal and Purkinje cells. The nervous fibers exhibited demyelinating lesions (showed small multifocal vacuole), and some axonal neuron atrophy gradually disappeared (formed large vacuole). Conclusions Apoptosis in brain tissue cells was mainly related to the propagation path and cytotropism of CDV. The apoptosis of astrocytes, oligodendrocytes, and some neurons may play a significant role in the demyelinating pathogenesis in dogs with acute canine distemper. A lot of diverse nervous signs shown in the clinic may be related to different neuron apoptosis.

Previously, we had cross-sectionally explored the characteristics of T cell receptor (TCR) repertoires from occupational medicamentosa-like dermatitis due to trichloroethylene (OMDT) patients, now we further analyzed the dynamic features of OMDT TCR repertoires. Peripheral blood TCR β-chain complementarity-determining region 3 (CDR3) genes were detected with the high throughput sequencing in 24 OMDT cases in their acute, chronic and recovery stages, respectively, and in 24 trichloroethylene-exposed healthy controls. The TCR repertoire diversity, TRBV/TRBD/TRBJ gene usage and combination, frequencies of CDR3 nucleotide (nt) and amino acid (aa) sequences in the cases in different stages and in the controls were analyzed. TRBV6-4 and TRBV7-9 frequencies significantly differed between the cases and controls (both P –4). TRBV6-4 combination with TRBJ2-1, TRBJ2-2, TRBJ2-3, and TRBJ2-6, and TRBV7-9 combination with TRBJ2-1 were associated with the stage by OMDT severity (all P P

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide and causing a global pandemic. Bat-origin RaTG13 is currently the most phylogenetically related virus. Here, we obtained the complex structure of RaTG13 receptor binding domain (RBD) with human ACE2 (hACE2), and further evaluated the binding of RaTG13 RBD to 24 additional ACE2 orthologs. By substituting residues in RaTG13 RBD with their counterparts in SARS-CoV-2 RBD, we found that residue 501, the major position found in VOCs 501Y.V1/V2/V3, plays a key role in determining the potential host range of RaTG13. We also found that SARS-CoV-2 could induce strong cross-reactive antibodies to RaTG13 and identified a SARS-CoV-2 MAb, CB6, that could cross-neutralize RaTG13 pseudovirus. These results elucidate the receptor binding and host-adaption mechanisms of RaTG13 and emphasize the importance of continuous surveillance of coronaviruses (CoVs) carried by animal reservoirs to prevent another spill-over of CoVs., Structural and molecular analysis of the receptor binding domain of RaTG13, a coronavirus closely related phylogenetically to SARS-CoV-2, bound to the human receptor ACE2 as well as ACE-2 orthologs in 24 other species provides a framework to understand its host range as well as the basis of antibody cross-reactivity between the two viruses.

This study aimed to assess the clinicopathological significance of serum levels and endometrium tissue expression of visfatin in polycystic ovary syndrome (PCOS) patients. A total of 80 PCOS patients and 80 matching controls were included in this study. We analyzed the relationship between the expression of visfatin in endometrium and clinicopathological characteristics in PCOS patients. The correlation between the expression of visfatin and p-Akt, Akt, p-ERK1/2, and ERK1/2 in PCOS tissues was evaluated as well. Visfatin expression in PCOS endometrial tissues were significantly higher than those in controls (

Objective Despite evidence that estrogens and insulin are involved in the development and progression of many cancers, their synergistic role in endometrial carcinoma (EC) has not been analyzed yet. Methods Here, we investigated how estrogens act synergistically with insulin to promote EC progression. Cell growth in vitro and in vivo, effects of estradiol and insulin on apoptosis and cell cycle distribution, and expression and activation of estrogen receptor (ER), insulin receptor (InsR), and key proteins in the PI3K and MAPK pathways were examined after combined stimulation with estradiol and insulin. Results Compared to EC cells treated with estradiol or insulin alone, those treated with both estradiol and insulin exhibited stronger stimulation. Estradiol significantly induced phosphorylation of InsR-β and IRS-1, whereas insulin significantly induced phosphorylation of ER-α. In addition, treatment with both insulin and estradiol together significantly increased the expression and phosphorylation of Akt, MAPK, and ERK. Notably, InsR-β inhibition had a limited effect on estradiol-dependent proliferation, cell cycle, and apoptosis, whereas ER-α inhibition had a limited insulin-dependent effect, in EC cell lines. Insulin and estradiol individually and synergistically promoted EC xenograft growth in mice. Conclusions Estrogen and insulin play synergistic roles in EC carcinogenesis and progression by activating InsR-β and ER-α, promoting a crosstalk between them, and thereby resulting in the activation of downstream PI3K/Akt and MAPK/ERK signaling pathways.

This study examined the temporal physiological and molecular events following the treatment of the liver with a tissue ablation modality that combined electroporation with electrolysis (E2). Rat liver was treated with an E2 waveform and the tissue examined, 1 h, 3 h, 6 h and 24 h with: HE, Masson Trichrome, TUNEL stains and Western blot. HE and TUNEL stains have shown that cell death began to be evident 3 h and hepatocyte regeneration was seen 24 h after treatment. HE and Masson trichrome have shown that the extracellular matrix and the large lumens, appeared intact after E2. Western blot has shown the following molecular events after E2: cleaved caspase 3-downgraded at 1 h, upgraded at 24 h (apoptosis); cleaved Caspase 1 and cleaved GSDMD-upgraded at 6 h (pyroptosis), RIP3-upgraded at 1 h, MLKL-upgraded at 3 h (necroptosis). The mechanism of cell death was possible initiated by necroptosis pathway. Pyroptosis pathway was also activated. The observation that cell death from E2 was by programed necrosis and the details on the temporal molecular pathways, may have value for the recent attempt to combine electroporation mediated ablation with immunological treatment, by demonstrating that the cell death from E2 involves an inflammatory response and by providing data that could be used to design the optimal timing for the injection of immunological adjuvants.

ObjectivesAccumulated evidence implies that night shift work may trigger liver dysfunction. Non-alcoholic fatty liver (NAFL) is suggested to be a necessary mediator in this process. This study aimed to examine the relationship between night shift work and elevated level of alanine transaminase (e-ALT) of workers and investigate the potential mediation effect of NAFL.MethodsThis study included all male workers from the baseline survey of a cohort of night shift workers. Information on demographics, lifestyle and lifetime working schedule was collected by face-to-face interview. Liver sonography was used to identify NAFL cases. Serum ALT level was detected by an automatic biochemical analyser. e-ALT was defined as ALT >40 U/L. Logistic regression models were used to evaluate ORs, and mediation analysis was employed to examine the mediation effect.ResultsAmong 4740 male workers, 39.5% were night shift workers. Night shift workers had an increased risk of e-ALT (OR, 1.19, 95% CI 1.00 to 1.42). With the increase in night shift years, the OR of e-ALT increased from 1.03 (95% CI 0.77 to 1.36) to 1.60 (95% CI 1.08 to 2.39) among workers without NAFL. A similar trend was not found among workers with NAFL. In addition, no significant mediation effect of NAFL in the association between night shift work and e-ALT was found.ConclusionsNight shift work is positively associated with abnormal liver function, in particular among workers without NAFL. Shift work involving circadian disruption is likely to exert a direct effect on liver dysfunction rather than rely on the mediation effect of NAFL.

Chronic benzene poisoning (CBP) is one of the most common chronic occupational poisoning which is associated with mitochondrial oxidative damage, and lead to increasing risk of respiratory diseases such as lung cancer. Cytochrome c oxidase subunit I (COI) is one of the key enzymes that plays an important role in oxidative damage regulation by eliminating reactive oxygen species (ROS). This study investigated the relationship between COI gene variants and the risk of CBP.We investigated 44 non-smoking patients who were diagnosed with CBP and 57 unexposed non-smoking controls between the ages of 23 and 60 with their background including work experience, lifestyle and medical records. Peripheral blood (2 mL) was collected in EDTA tube and the platelet was purified from the collected blood. Variants of COI were analyzed by PCR and sequencing. Multivariable linear regression analysis was used to assess the association between CBP exposure and variants.The frequency of the mitochondrial DNA (mtDNA) T6392C, G6962 variants were 10, 7 out of 44 CBP group patients, which was higher when compared to that of 4, 2 out of 57 in the control group, suggesting these variants could be the risk factor for CBP [odds ratio (OR) 3.897, 95% CI: 1.131-13.425, P=0.023; OR 5.203, 95% CI: 1.024-26.442, P=0.034]. There was a significant difference (P0.05) of COI variants, including T6392C and G6962A, in platelet mtDNA between patients and control samples. Meanwhile, the frequency of the mtDNA C7196A variant were 13 out of 44 control group, which was higher when compared to that of 2 of 57 in the CBP group patients, suggesting this variant could be the protective factor for CBP (OR 6.205, 95% CI: 1.320-29.162, P=0.010).Our study suggests that T6392C, G6962A and C7196A from platelet mtDNA variants play a significant role in the etiology of CBP and facilitate the development of molecular biomarker on CBP diagnosis.

N 6-methyladenosine (m6A) constitutes one of the most abundant internal RNA modifications and is critical for RNA metabolism and function. It has been previously reported that viral RNA contains internal m6A modifications; however, only recently the function of m6A modification in viral RNAs has been elucidated during infections of HIV, hepatitis C virus and Zika virus. In the present study, we found that enterovirus 71 (EV71) RNA undergoes m6A modification during viral infection, which alters the expression and localization of the methyltransferase and demethylase of m6A, and its binding proteins. Moreover, knockdown of m6A methyltransferase resulted in decreased EV71 replication, whereas knockdown of the demethylase had the opposite effect. Further study showed that the m6A binding proteins also participate in the regulation of viral replication. In particular, two m6A modification sites were identified in the viral genome, of which mutations resulted in decreased virus replication, suggesting that m6A modification plays an important role in EV71 replication. Notably, we found that METTL3 interacted with viral RNA-dependent RNA polymerase 3D and induced enhanced sumoylation and ubiquitination of the 3D polymerase that boosted viral replication. Taken together, our findings demonstrated that the host m6A modification complex interacts with viral proteins to modulate EV71 replication.

Background: To evaluate the effects of body weight loss on pregnancy and livebirth outcomes in young women with early-stage endometrial cancer (EC) and atypical hyperplasia (AH) with fertility-sparing therapy. Thus, improve the management of this patient group.Method: Patients with AH (n=36) and well-differentiated EC (n=8, FIGO stage IA) who achieved complete regression after conservative treatment were included in this retrospective study. A weight loss group (n=25) and a non-weight loss group (n=19) were divided; while subgroup analysis according to body mass index and stratification analysis according to weight loss proportion were performed to investigate the effect of weight loss on pregnancy and livebirth outcomes. A univariate and multivariate logistic regression analysis were undertaken to analysis the factors associated with pregnancy.Results: The mean body weight and body mass index at pretreatment of progestin and initiation of fertility treatment was 70.63±12.03 and 67.08±8.18 kg, respectively, and 27.06±4.44 and 25.73±3.15 kg/m2, respectively. 25 patients (56.82%) had weight loss; the median weight loss amount is 5.00kg (1.00-34.50), median weigh loss proportion was 6.70% (1.00-36.00) during median time interval of 12months (5.00-97.00). An impressive favorable pregnancy rate (65.91%) and live birth rate (50.00%) were achieved. The pregnancy and livebirth rate were meaningfully higher in the weight loss group than the non-weight loss group (88.00% vs.36.84%,P=0.000; 64.00% vs.31.58%,P=0.033, respectively); weight loss≥5% significantly increased pregnancy and live birth rate in patients with BMI≥25. The risk ratios of weight loss≥5% in multivariate logistic analysis for pregnancy was 0.096(0.010, 0.907).Conclusions: Weight loss could have a positive effect on pregnancy rates and seem to be useful for improving live birth rates in overweight or obese women with early-stage endometrial cancer and atypical hyperplasia during/after fertility-sparing therapy. weight loss≥5% was protective factors of pregnancy in fertility-sparing patients with early-stage endometrial cancer and atypical hyperplasia.

Avian reovirus (ARV) can induce many diseases as well as immunosuppression in chickens, severely endangering the poultry industry. Interferons (IFNs) play an antiviral role by inducing the expression of interferon-stimulated genes (ISGs). The effect of ARV infection on the expression of host ISGs is unclear. Specific-pathogen-free (SPF) chickens were infected with ARV strain S1133 in this study, and real time quantitative PCR was used to detect changes in the dynamic expression of IFNs and common ISGs in joints of SPF chickens. The results showed that the transcription levels of IFNA, IFNB, and several ISGs, including myxovirus resistance (MX), interferon-induced transmembrane protein 3 (IFITM3), protein kinase R (PKR), oligoadenylate synthase (OAS), interferon-induced protein with tetratricopeptide repeats 5 (IFIT5), interferon-stimulated gene 12 (ISG12), virus inhibitory protein (VIPERIN), interferon-alpha-inducible protein 6 (IFI6), and integrin-associated protein (CD47), were upregulated in joints on days 1–7 of infection (the levels of increase of MX, IFIT5, OAS, VIPERIN, ISG12, and IFI6 were the most significant, at hundreds-fold). In addition, the expression levels of the ISGs encoding zinc finger protein 313 (ZFP313), and DNA damage–inducible transcript 4 (DDIT4) increased suddenly on the 1st or 2nd day, then decreased to control levels. The ARV viral load in chicken joints rapidly increased after 1 day of viral challenge, and the viral load remained high within 6 days of viral challenge. The ARV viral load sharply decreased starting on day 7. These results indicate that in SPF chicken joints, many ISGs have mRNA expression patterns that are basically consistent with the viral load in joints. IFNA, IFNB, and the ISGs MX, IFITM3, PKR, OAS, IFIT5, ISG12, VIPERIN, IFI6, and CD47 play important roles in defending against ARV invasion, inhibiting ARV replication and proliferation, and promoting virus clearance. These results enrich our understanding of the innate immune response mechanisms of hosts against ARV infection and provide a theoretical basis for prevention and control of ARV infection.