IL-6/STAT3 axis is hijacked by GCRV to facilitate viral replication via suppressing type â IFN signaling.

Grass carp reovirus (GCRV) infections and hemorrhagic disease (GCHD) outbreaks are typically seasonally periodic and temperature-dependent, yet the molecular mechanism remains unclear. Herein, we depicted that temperature-dependent IL-6/STAT3 axis was exploited by GCRV to facilitate viral replication via suppressing type Ⅰ IFN signaling. Combined multi-omics analysis and qPCR identified IL-6, STAT3, and IRF3 as potential effector molecules mediating GCRV infection. Deploying GCRV challenge at 18 °C and 28 °C as models of resistant and permissive infections and switched to the corresponding temperatures as temperature stress models, we illustrated that IL-6 and STAT3 expression, genome level of GCRV, and phosphorylation of STAT3 were temperature dependent and regulated by temperature stress. Further research revealed that activating IL-6/STAT3 axis enhanced GCRV replication and suppressed the expression of IFNs, whereas blocking the axis impaired viral replication. Mechanistically, grass carp STAT3 inhibited IRF3 nuclear translocation via interacting with it, thus down-regulating IFNs expression, restraining transcriptional activation of the IFN promoter, and facilitating GCRV replication. Overall, our work sheds light on an immune evasion mechanism whereby GCRV facilitates viral replication by hijacking IL-6/STAT3 axis to down-regulate IFNs expression, thus providing a valuable reference for targeted prevention and therapy of GCRV.

Corrigendum to "Prognostic Factors for COVID-19 Hospitalized Patients with Preexisting Type 2 Diabetes".

[This corrects the article DOI: 10.1155/2022/9322332.].

Prognostic Factors for COVID-19 Hospitalized Patients with Preexisting Type 2 Diabetes.

BACKGROUND: Type 2 diabetes (T2D) as a worldwide chronic disease combined with the COVID-19 pandemic prompts the need for improving the management of hospitalized COVID-19 patients with preexisting T2D to reduce complications and the risk of death. This study aimed to identify clinical factors associated with COVID-19 outcomes specifically targeted at T2D patients and build an individualized risk prediction nomogram for risk stratification and early clinical intervention to reduce mortality. METHODS: In this retrospective study, the clinical characteristics of 382 confirmed COVID-19 patients, consisting of 108 with and 274 without preexisting T2D, from January 8 to March 7, 2020, in Tianyou Hospital in Wuhan, China, were collected and analyzed. Univariate and multivariate Cox regression models were performed to identify specific clinical factors associated with mortality of COVID-19 patients with T2D. An individualized risk prediction nomogram was developed and evaluated by discrimination and calibration. RESULTS: Nearly 15% (16/108) of hospitalized COVID-19 patients with T2D died. Twelve risk factors predictive of mortality were identified. Older age (HR = 1.076, 95% CI = 1.014-1.143, p=0.016), elevated glucose level (HR = 1.153, 95% CI = 1.038-1.28, p=0.0079), increased serum amyloid A (SAA) (HR = 1.007, 95% CI = 1.001-1.014, p=0.022), diabetes treatment with only oral diabetes medication (HR = 0.152, 95%CI = 0.032-0.73, p=0.0036), and oral medication plus insulin (HR = 0.095, 95%CI = 0.019-0.462, p=0.019) were independent prognostic factors. A nomogram based on these prognostic factors was built for early prediction of 7-day, 14-day, and 21-day survival of diabetes patients. High concordance index (C-index) was achieved, and the calibration curves showed the model had good prediction ability within three weeks of COVID-19 onset. CONCLUSIONS: By incorporating specific prognostic factors, this study provided a user-friendly graphical risk prediction tool for clinicians to quickly identify high-risk T2D patients hospitalized for COVID-19.

Metformin attenuates PD-L1 expression through activating Hippo signaling pathway in colorectal cancer cells.

Programmed cell death ligand 1 (PD-L1) is a key suppressor of the cytotoxic immune response. In colorectal carcinoma (CRC), PD-L1 expression results in immune escape and poor prognosis. Extensive researches suggested that metformin had a potential efficacy of enhancing anti-tumor immune response in different types of cancer. However, the detail mechanisms underlying the efficacy in CRC are unclear. Here, we showed that metformin decreases PD-L1 and YAP1 expression in vitro and in vivo. After silencing or inhibiting YAP1 expression by Verteporfin (VP), the inhibitor of YAP1, the expression of PD-L1 were decreased in protein level in CRC cells. Furthermore, metformin directly phosphorylated YAP1 and restricted YAP1 to entry in the nucleus, so that PD-L1 was reduced via western blot and immunofluorescence assays in SW480 and HCT116 cells. Finally, subcutaneous xenotransplanted tumor models of HCT116 cells were established in BALB/c nude mice. Compared with the control group, PD-L1 and YAP1 expressions in tumor tissues, detected by immunohistochemistry, were reduced in the group of metformin treatment. These findings illuminate a new regulatory mechanism, metformin activates Hippo signaling pathway to regulate PD-L1 expression and suggests that metformin has the possibility to increase the efficacy of immunotherapy in human CRC.

Delayed cognitive deficits can be alleviated by calcium antagonist nimodipine by downregulation of apoptosis following whole brain radiotherapy.

Radiation therapy is important for the comprehensive treatment of intracranial tumors. However, the molecular mechanisms underlying the pathogenesis of delayed cognitive dysfunction are not well-defined and effective treatments or prevention measures remain insufficient. In the present study, 60 adult male Wistar rats were randomly divided into three groups, which included a control, whole brain radiotherapy (WBRT) (single dose of 30 Gy of WBRT) and nimodipine (single dose of 30 Gy of WBRT followed by nimodipine injection intraperitoneally) groups. The rats were sacrificed 7 days or 3 months following irradiation. At 3 months, the Morris water maze test was used to assess spatial learning and memory function in rats. The results demonstrated that the WBRT group demonstrated a significantly impaired cognitive performance, decreased numbers of hippocampal Cornu Ammonis (CA)1 neurons and upregulated expression of caspase-3 in the dentate gyrus compared with those in the control and nimodipine groups. Reverse transcription-quantitative polymerase chain reaction analysis demonstrated that the WBRT group exhibited increased ratio of B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax)/Bcl-2 compared with that in control and nimodipine groups on day 7 following irradiation. However, the WBRT group exhibited decreased levels of brain-derived neurotrophic factor (BDNF) compared with that in control and nimodipine groups at 3 months following brain irradiation. The levels of growth-associated protein 43 and amyloid precursor protein between the nimodipine group and WBRT group were not statistically significant. The present study demonstrated that neuron apoptosis may lead to delayed cognitive deficits in the hippocampus, in response to radiotherapy. The cognitive impairment may be alleviated in response to a calcium antagonist nimodipine. The molecular mechanisms involved in nimodipine-mediated protection against cognitive decline may involve the regulation of Bax/Bcl-2 and BDNF in the hippocampus.

[Influence of registration based on different reference markers on the displacement of the geometry consisted of all clips in the cavity for external-beam partial breast irradiation at moderate deep inspiration breath holding].

OBJECTIVE: To investigate the influence of registration based on different reference markers on the displacement of the geometry consisted of all clips in the cavity for external-beam partial breast irradiation at moderate deep inspiration breath holding assisted by active breathing control device. METHODS: Twenty-seven early stage breast cancer patients feasible for external beam partial breast irradiation (EB-PBI) were selected. The patients undertaken three-dimensional computed tomography (3DCT) simulation scan at moderate deep inspiration breath holding (mDIBH) assisted by active breathing control device, and two sets of mDIBH CT images were got and transferred to the Pinnacle 3 planning system. All of the silver clips were delineated and a geometry consisted of all clips were generated. On the account of automatic registration of mDIBH CT images, manual registration was carried out based separately on the topside clip in the cavity, the labeled skin at anterior surface of the cavity at central level and the metal mark on the body surface near the cavity, then the displacements of center of the geometry in left-right (LR), anterior-posterior (AP) and superior-inferior (SI) directions based separately on the three registrations were measured. RESULTS: The displacements of center of the geometry in LR, AP and SI directions based on registration of the clips, the labeled skin and the metal mark were (0.61 ± 0.62)mm vs. (1.11 ± 1.21)mm vs. (1.31 ± 1.55)mm, (0.63 ± 0.59)mm vs. (0.92 ± 0.93)mm vs. (1.19 ± 1.24)mm and (0.91 ± 0.96)mm vs. (2.13 ± 2.12)mm vs. (1.93 ± 1.55)mm, respectively. Compared the displacements of center of the geometry in the same direction between the three registrations, significant differences were found only in SI direction between clip registration and skin registration, clip registration and mark registration (t = 5.045, 7.210 and P = 0.025, 0.007) . Compared the displacements of center of the geometry between three dimensional directions for each reference registration, there was no significant difference (all P > 0.05). CONCLUSIONS: When EB-PBI is carried out in state of mDIBH, measurement of the intrafraction displacement based on registration of the clip in the cavity is a reasonable selection. Otherwise, excessive margin enlargement of PTV in SI direction will be generated if the regional skin or metal mark is selected as registration reference.

Effects of anisodamine on the expressions of vascular endothelial growth factor and intercellular adhesion molecule 1 in experimental infusion phlebitis.

BACKGROUND: Infusion phlebitis is the most common side effect of clinical intravenous drug therapy and several clinical studies have demonstrated that anisodamine can effectively prevent the occurrence of infusion phlebitis. This study was designed to investigate effects of anisodamine on the expressions of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule 1 (ICAM-1) in a rabbit model of infusion phlebitis and to analyze the mechanisms of anisodamine effect on the prevention and treatment of experimental infusion phlebitis. METHODS: Twenty-four specific pathogen-free male Japanese white rabbits were randomly assigned to the control group, the model group, the magnesium sulfate group and the anisodamine group. The rabbit model of infusion phlebitis, induced by intravenous administration, was established and expressions of VEGF and ICAM-1 were determined and contrasted with the control group treated with normal saline. We evaluated expression by histopathology, immunohistochemistry, reverse transcription-polymerase chain reaction, and Western blotting assay. RESULTS: Pathohistological changes of the model group were observed, such as loss of venous endothelial cells, inflammatory cell infiltration, edema and thrombus. The magnesium sulfate group and the anisodamine group showed significant protective effects on vascular congestion, inflammatory cell infiltration, proliferation, swelling of endothelium and perivascular hemorrhage. The model group showed the highest expressions of VEGF and ICAM-1 of the four groups (P < 0.01). On the contrary, anisodamine alleviated the inflammatory damage by significantly reducing the expressions of VEGF and ICAM-1 compared with the model group (P < 0.01). There was no significant difference in the expressions of VEGF and ICAM-1 between the magnesium sulfate group and the anisodamine group (P > 0.05). CONCLUSION: Anisodamine alleviates inflammatory damage by significantly reducing the expressions of VEGF and ICAM-1, and shows significant protective effects in an animal model of infusion phlebitis.

[Comparison of overlap ratios of the target volume in different respiratory states with active breathing control for external-beam partial breast irradiation].

OBJECTIVE: To explore the overlap ratio of the target volume in different respiratory statuses of active breath control (ABC) and their differences during external-beam partial breast irradiation (EB-PBI), and from the perspective of target volume overlap to determine the influence of the ABC-assisted breathing condition on intra-fractional target movement of EB-PBI. METHODS: The patients, who received breast-conserving surgery with silver clips marked at the margins of the cavity and were suitable for EB-PBI, were immobilized on the breast bracket to undertake CT simulation assisted by ABC device, six sets of CT simulation images including two sets of image in state of moderate deep inspiration breathing control (mDIBH), two sets of images in state of free breath (FB) and two sets of images in state of deep expiration breathing control (DEBH) were obtained. The six sets of images were transferred to Pinnacle(3) treatment planning system (TPS), then automatic fusion and registration between two sets of mDIBH images, two sets of FB images, two sets of DEBH images and mDIBH image and DEBH image were achieved separately. Thereafter, the overlap ratios of GTV with GTV, CTV with CTV, PTV with PTV were calculated by the Pinnacle(3) TPS. The differences between the overlap ratios of the three kinds of targets in the same registered image and the difference between the overlap ratios of the same kind of target in the different registered images were statistically analyzed using statistical package of SPSS 11.5. RESULTS: Based on mDIBH/mDIBH registration, the overlap ratios of GTV/GTV, CTV/CTV and PTV/PTV were (83.54 ± 11.41)%, (93.00 ± 6.49)%, and (95.26 ± 4.90)%, respectively, and the differences of the overlap ratios between GTV/GTV and CTV/CTV, GTV/GTV and PTV/PTV were all statistically significant (P < 0.05), but statistically not significant between CTV/CTV and PTV/PTV (P > 0.05). Based on FB/FB registration, the overlap ratios of GTV/GTV, CTV/CTV and PTV/PTV were (72.55 ± 29.10)%, (89.36 ± 9.53)% and (92.47 ± 7.25)%, respectively, and the differences of the overlap ratios between GTV/GTV and CTV/CTV, CTV/CTV and PTV/PTV were all not statistically significant (P > 0.05), but statistically significant between GTV/GTV and PTV/PTV (P < 0.05). Based on DEBH/DEBH registration, the overlap ratios of GTV/GTV, CTV/CTV and PTV/PTV were (79.48 ± 22.31)%, (92.83 ± 6.77)% and (95.05 ± 4.81)%, respectively, and the differences of the overlap ratios between GTV/GTV and CTV/CTV (P = 0.000), CTV/CTV and PTV/PTV (P = 0.037), GTV/GTV and PTV/PTV (P = 0.000) were statistically all significant (P = 0.000). The differences of the overlap ratios of GTV/GTV, CTV/CTV, and PTV/PTV (P = 0.000) between mDIBH/mDIBH and DEBH/DEBH, mDIBH/mDIBH and FB/FB, FB/FB and DEBH/DEBH were all statistically significant (P = 0.000), and not statistically significant between mDIBH/mDIBH and mDIBH/DEBH, FB/FB and mDIBH/DEBH. CONCLUSIONS: During the delivering of EB-PBI assisted by ABC, the intra-fractional overlap ratios of the target volume between the same breathing state is increasing in the order of GTV/GTV → CTV/CTV → PTV/PTV. The difference of the overlap ratios of the target volumes between mDIBH and mDIBH, FB and FB, DEBH and DEBH is not significant, and the overlap ratios of PTV/PTV in the three breathing statuses of mDIBH, FB and DEBH reaches a higher level. Therefore, from the perspective of target volume overlap, if the setup error is corrected online before delivering, the necessity of breathing control during delivering of EB-PBI is worthy discussing.

A case of 48,XXYY syndrome detected prenatally by QF-PCR.

We report on the prenatal diagnosis and genetic analysis of a 48,XXYY fetus. A 28-year-old woman was referred for amniocentesis at 18 weeks' gestation because of advanced paternal age. Quantitative-fluorescence polymerase chain reaction (QF-PCR) with small tandem repeat (STR) markers rapidly detected the sex chromosome polysomy from amniotic fluid cells. This abnormality appeared to arise from successive non-disjunction during paternal meiosis I and meiosis II. Cytogenetic analysis revealed a karyotype of 48,XXYY. This case adds to the evidence that an age-related increase in sex chromosomal aneuploidies occurs in sperm. This case also shows that QF-PCR assays can provide rapid prenatal diagnosis of numerical sex chromosome aneuploidy.

[MMP-2/TIMP-2 expression in the trophoblasts of patients with gestational trophoblastic disease].

OBJECTIVE: To explore the role of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of MMP-2 (TIMP-2) in the pathogenesis, development and prognosis of gestational trophoblastic disease (GTD). METHODS: In situ hybridization and immunohistochemistry were utilized for MMP-2/TIMP-2 mRNA and protein detection in normal chorion of women with early gestation, hydatidiform mole, invasive mole, or choricarcinoma. RESULTS: The results revealed that specific staining for mRNA and protein of MMP-2 and the expression of TIMP-2 was reduced in normal chorion of early gestation. In GTD ranging from hydatidiform mole, invasive mole to choricarcinoma, MMP-2 expression tended to increase while TIMP-2 expression underwent an invert change. The positivity rate of MMP-2 and TIMP-2 in gestational trophoblastic tumor group was higher than that of the normal chorion of early gestation group and hydatiform mole group (P<0.05 and P<0.001, respectively). CONCLUSION: A disrupted balance between the activation and inhibition of MMP-2 plays a critical role in the pathogenesis, progression and metastasis of GTD.