Objective To elucidate the correlation between specific nucleocapsid(N)protein mutant of the SARS-CoV-2 B.1.1.7 variant and clinical stratification in COVID-19 patients,revealing their impact on N protein liquid-liquid phase separation(LLPS)and host innate immune response.Methods Based on whole-genome sequencing data of the SARS-CoV-2 B.1.1.7 lineage from the GISAID database,non-synonymous mutation sites significantly associated with mild/severe clinical phenotypes were screened.For high-frequency N protein mutant,IFN-β promoter transcriptional activity was quantitatively measured using a dual-luciferase reporter system.qPCR was used to detect the mRNA expression levels of interferon(IFN)-β,interleukin(IL)-6 and tumor necrosis factor(TNF)-α.LLPS characteristics were observed by confocal microscopy.The ubiquitination status of host MAVS was detected by Western blot assay.Results A total of 17 640 non-synonymous mutation sites were identified,among which 65 were associated with mild cases and 20 were related to severe cases,with a mutation frequency>1%.The N protein mutation sites associated with severe cases were D3L,M234I and R203K-G204R-T205I.N protein and the mutants NM234I,NR203K-G204R-T205I inhibited the promoter activity of IFN-β(P＜0.05).Compared to the wild type N protein,NR203K-G204R-T205I mutation significantly reduced the mRNA levels of IFN-β,IL-6 and TNF-α(P＜0.05),and altered the phase separation state by dispersing the formation of LLPS condensates.However,N mutant did not affect the ubiquitination modification of host MAVS.Conclusion N protein mutants of the SARS-CoV-2 B.1.1.7 variant can influence the clinical prognosis of COVID-19 patients by altering LLPS status and suppressing the innate immune responses.These finding provides a theoretical basis for the design of antiviral drugs targeting the N protein.

Objective To elucidate the correlation between specific nucleocapsid(N)protein mutant of the SARS-CoV-2 B.1.1.7 variant and clinical stratification in COVID-19 patients,revealing their impact on N protein liquid-liquid phase separation(LLPS)and host innate immune response.Methods Based on whole-genome sequencing data of the SARS-CoV-2 B.1.1.7 lineage from the GISAID database,non-synonymous mutation sites significantly associated with mild/severe clinical phenotypes were screened.For high-frequency N protein mutant,IFN-β promoter transcriptional activity was quantitatively measured using a dual-luciferase reporter system.qPCR was used to detect the mRNA expression levels of interferon(IFN)-β,interleukin(IL)-6 and tumor necrosis factor(TNF)-α.LLPS characteristics were observed by confocal microscopy.The ubiquitination status of host MAVS was detected by Western blot assay.Results A total of 17 640 non-synonymous mutation sites were identified,among which 65 were associated with mild cases and 20 were related to severe cases,with a mutation frequency>1%.The N protein mutation sites associated with severe cases were D3L,M234I and R203K-G204R-T205I.N protein and the mutants NM234I,NR203K-G204R-T205I inhibited the promoter activity of IFN-β(P＜0.05).Compared to the wild type N protein,NR203K-G204R-T205I mutation significantly reduced the mRNA levels of IFN-β,IL-6 and TNF-α(P＜0.05),and altered the phase separation state by dispersing the formation of LLPS condensates.However,N mutant did not affect the ubiquitination modification of host MAVS.Conclusion N protein mutants of the SARS-CoV-2 B.1.1.7 variant can influence the clinical prognosis of COVID-19 patients by altering LLPS status and suppressing the innate immune responses.These finding provides a theoretical basis for the design of antiviral drugs targeting the N protein.

Humans ; Soft Tissue Neoplasms ; Mesenchymoma/diagnostic imaging* ; Paraneoplastic Syndromes/pathology*

Objective:To explore the risk factors of poor renal prognosis in patients with coronary artery bypass surgery (CABG)-associated acute kidney injury (AKI), and establish a preliminary clinical risk prediction model for chronic kidney disease (CKD) progression in CABG-associated AKI patients, and evaluate its predictive efficacy.Methods:It was a retrospective, observational cohort study. The study subjects were patients who underwent CABG at Central China Fuwai Hospital from January 1, 2018 to December 31, 2020, with a baseline estimated glomerular filtration rate (eGFR)>60 ml·min -1·(1.73 m 2) -1 and postoperative complication of AKI. The patients were followed up for 90 days after discharge from hospital. The endpoint event was defined as progression to CKD after 90 days of the occurrence of CABG-associated AKI. The patients were divided into CKD group and non-CKD group based on whether they experienced endpoint events. The baseline clinical data were compared between the two groups. The logistic regression model was used to analyze the risk factors of endpoint event. The receiver-operating characteristic curve was drawn to evaluate the performance of the clinical risk prediction model for predicting poor renal prognosis in CABG-associated AKI patients. Results:A total of 124 CABG-associated AKI patients were enrolled in the study, including 95 males and 29 females, aged (62.57±9.61) years old. Thirty-eight patients (30.8%) progressed to new-onset CKD 90 days after CABG-associated AKI. Compared with non-CKD group, CKD group had lower preoperative hemoglobin ( t=2.778, P=0.006) and baseline eGFR ( t=3.603, P<0.001), higher proportion of women ( χ2=10.714, P=0.001), and higher preoperative blood NT-proBNP ( Z=-2.150, P=0.030) and discharged serum creatinine ( Z=-5.290, P<0.001). The multivariate logistic regression analysis results revealed that female ( OR=5.179, 95% CI 1.535-17.477, P=0.008), high preoperative blood NT-proBNP ( OR=1.049, 95% CI 1.004-1.095, P=0.032), low baseline eGFR ( OR=0.928, 95% CI 0.889-0.968, P=0.001), and high serum creatinine at discharge ( OR=1.019, 95% CI 1.009-1.029, P<0.001) were independent influencing factors of CABG-associated AKI to CKD. The clinical risk prediction model including female, preoperative blood NT-proBNP, preoperative baseline eGFR, and serum creatinine at discharge produced a moderate performance for predicting CABG-associated AKI to CKD ( AUC=0.872, 95% CI 0.806-0.939, P<0.001). Conclusion:Patients with CABG-associated AKI are at high risks for new-onset CKD. Female, preoperative high NT-proBNP, preoperative low baseline eGFR and high serum creatinine at discharge can help identify patients with a high risk of CABG-associated AKI to CKD.

Glioblastoma (GBM) is a highly aggressive and lethal brain tumor with an immunosuppressive tumor microenvironment (TME). In this environment, myeloid cells, such as myeloid-derived suppressor cells (MDSCs), play a pivotal role in suppressing antitumor immunity. Lipometabolism is closely related to the function of myeloid cells. Here, our study reports that acetyl-CoA acetyltransferase 1 (ACAT1), the key enzyme of fatty acid oxidation (FAO) and ketogenesis, is significantly downregulated in the MDSCs infiltrated in GBM patients. To investigate the effects of ACAT1 on myeloid cells, we generated mice with myeloid-specific (LyzM-cre) depletion of ACAT1. The results show that these mice exhibited a remarkable accumulation of MDSCs and increased tumor progression both ectopically and orthotopically. The mechanism behind this effect is elevated secretion of C-X-C motif ligand 1 (CXCL1) of macrophages (Mφ). Overall, our findings demonstrate that ACAT1 could serve as a promising drug target for GBM by regulating the function of MDSCs in the TME.

Objective To assess the prognostic value of KRAS mutation in patients with advanced primary liver cancer treated with transcatheter arterial chemoembolization (TACE). Methods Ninety-seven patients with advanced primary liver cancer who received TACE treatment in The Third People's hospital from April 2017 to May 2020 were included. The mutation status of KRAS was detected, and its relationship with the prognosis of TACE was investigated. The t -test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. Survival analysis was performed using Kaplan-Meier survival curve and compared using Log-rank test. Cox regression analysis was performed to identify the prognostic factors. Results Among 97 patients with advanced liver cancer, KRAS mutations were detected in 34 patients (35.05%), including 21 patients with codon 12 mutation (61.76%) and 13 patients with codon 13 mutation (38.24%). KRAS mutation was associated with liver cirrhosis, intrahepatic metastasis and the number of tumors ( χ 2 =0.035, 3.965, and 6.593, all P < 0.05). Survival analysis showed that the progression free survival and overall survival were significantly longer in KRAS wild-type patients than in KRAS mutant patients ( χ 2 =4.465 and 4.280, all P < 0.05). Multivariate Cox analysis revealed that KRAS mutation, intrahepatic metastasis, number of tumors and BCLC stage were important factors affecting the overall survival and prognosis of patients (all P < 0.05). Conclusion KRAS mutation is common in patients with advanced primary liver cancer and is closely associated with a poor prognosis after TACE. It may become a potential indicator of clinical prognosis.

Objective By using CT-guided percutaneous inoculation with "external pre-loaded and traced one-step implantation technique" to establish VX2 liver and kidney tumor model in experimental rabbits, and to evaluate the effectiveness and convenience of this technique. Methods By using a disposal 16G thoracic puncture needle (length of 8. 5 cm) and a self-made 18 G puncture probe, inoculation of tumor cells was performed in experimental rabbits. Retrograde filling of gelatin sponge strip, which was infiltrated with 0. 3 mL contrast medium, into the thoracic puncture needle lumen was performed, which was followed by retrograde loading of 1X1X3 mm3 VX2 tissue block, and the pre-assembly in vitro was completed. Under CT guidance, percutaneous puncturing of the target organ of experimental rabbit with pre - assembly probe was conducted, then, the end of the needle was inserted into the probe to push out the tumor tissue and gelatin sponge when it was verified that the needle tip reached the desired target point. After 30 seconds, the needle and probe were pulled out slowly and integrally, and the puncture point was manually pressed for 60 seconds. CT scan was performed to check the location of high-density gelatin sponge. The tumor inoculation time was recorded. Contrast - enhanced CT scan was employed at 2, 3 and 4 weeks after inoculation to validate the modeling results. Results Immediate postoperative CT scan showed that the inoculation site was manifested as nodular high density shadow in the liver or kidney parenchyma. Liver tumor modeling was used in 10 rabbits, and tumor formation was obtained in all 10 rabbits (100％, 10/10), and the mean time used for inoculation was 4. 3 minutes. Kidney tumor modeling was adopted in 10 rabbits, and tumor formation was obtained in 9 rabbits (90％, 9/10), and the mean time used for inoculation was 4. 9 minutes. The inoculated tumor was characterized by a single lesion in all experimental rabbits, the location of the tumor was basically the same as that of the high density gelatin sponge. Check-up contrast-enhanced CT scan showed that the liver implanted tumor had peripheral enhancement, which presented as ring - shaped enhancement on DSA. The kidney implanted tumor was manifested as renal parenchyma staining defect on both contrast - enhanced CT scan and DSA. After 3-4 weeks of inoculation, the implanted tumors grew rapidly. Conclusion For the establishment of rabbit VX2 liver and kidney tumor models, CT - guided percutaneous inoculation with "external pre-loaded and traced one-step implantation technique" is very effective. This technique has several advantages, such as convenient and time-saving, postoperative immediate confirmation of the inoculation site, higher tumor formation rate, solitary tumor formation, etc. (J Intervent Radiol, 2018, 27:252-256)

OBJECTIVETo perform a prospective,multicenter,open,randomized study to determine a treatment regimen for treatment-naive patients with refractory chronic hepatitis C (RHC) using the predictive value (PV) of early virological response (EVR).

METHODSA total of 438 patients from 18 hospitals were recruited between December 2008 and December 2010 and administered peg-interferon/ribavirin treatment for 12 weeks. Patients who achieved complete EVR (cEVR) were assigned to group A for a 48-week course of treatment, while patients without cEVR were randomly allocated to either group B 1 for a 72-week course of treatment or to group B2 for a 96-week course of treatment. Serum hepatitis C virus RNA levels at baseline,treatment weeks 4, 12 and 24, end of treatment, and post-treatment week 24 were measured and used to evaluate the efficiency of therapy.

RESULTSThe overall sustained virological response (SVR) rate was 85.1%. In all, 91.0% of patients achieved cEVR and were assigned to group A, which had an SVR rate of 90.8%. There was no statistically significant difference in the SVR rates of groups B1 and B2 (29.4% vs. 25.0%, P more than 0.05). The positive PV of rapid virological response (RVR), cEVR and delayed virological response (DVR) for SVR was 93.4%, 90.8% and 77.8% respectively, and the negative PV of RVR, EVR and DVR for SVR was 28.0%, 93.3% and 100% respectively. Overall, 66.9% of the patients experienced adverse events (AEs), but only 1.9% of patients experienced sevcre AEs.

CONCLUSIONThe majority of Chinese RHC treatmentna(i)ve patients (91.0%) can achieve cEVR and a high SVR rate with a low rate of severe AEs using the cEVR guided personal treatment regimen.

Antiviral Agents ; Asian Continental Ancestry Group ; Drug Therapy, Combination ; Hepatitis C, Chronic ; Humans ; Ribavirin

BACKGROUNDInfancy is a critical period of growth and physiological development, in which breast milk is the best source of nutrients. Compared to western countries, research on breast milk of Chinese population are limited. Thus, it is necessary to measure breast milk energy and macronutrient concentrations of healthy urban Chinese mothers at different lactation stages, to expand the database of milk composition of Chinese population, and to examine whether dietary or other maternal factors can affect the levels of macronutrients in breast milk.

METHODSBreast milk of full expression of one side breast from 436 urban Chinese lactating mothers at 5-11 days, 12-30 days, 31-60 days, 61-120 days, and 121-240 days postpartum was obtained at 9:00 a.m. to 11:00 a.m. Total energy, lactose, protein, and fat contents were measured. 24-hour dietary recall was surveyed, and maternal nutrient intakes were analyzed.

RESULTSMilk composition changed over the course of lactation and large individual variations were documented. The concentrations were 61.3 kcal/dl for total energy, 7.1 g/dl for lactose, 0.9 g/dl for protein, and 3.4 g/dl for fat in mature milk. Stage of lactation was a strong factor affecting milk composition. Minimal evidence was found for associations between maternal current dietary intake and milk macronutrient concentration, consistently with prior research. Maternal body mass index (BMI) was positively associated with milk fat content, to a greater extent than did dietary intake. All other maternal characteristics were not significant for milk composition.

CONCLUSIONThese findings suggest that milk composition is generally weakly associated with maternal factors except for stage of lactation, and is likely to be more susceptible to long-term maternal nutritional status than short-term dietary fluctuation.

Adolescent ; Adult ; Body Mass Index ; Female ; Humans ; Lactation ; physiology ; Middle Aged ; Milk, Human ; chemistry ; Mothers ; Young Adult

Objective To probe the value of multi-slice spiral CT angiography (MSCTA) in decreasing surgical trauma of the cranial base meningioma. Methods Thirty-two patients with cranial base meningioma were examined preoperatively with MSCTA to observe the shape and the relationship with the adjacent vessels and the skull base. Three-dimensional images were reconstructed to imitate the approach of operation and compared with surgical findings. Meanwhile, 22 patients withnot MSCTA were selected randomly as control group. The amount of blood transfusion and the occurrence rate of complications were compared between the two groups. Results MSCTA depiceted clear three dimensional images of the meningioma and the relationship with the adjacent vessels and the skull base, corresponded very well to the surgery. By imitating the operation, all patients were designed the incision size of bone appropriately, the vessels of peritumoral were kept off effectively and the risk of the embedded vascular were assessed accurately. The conventional surgical approach and method were changed in 9 patients, 4 formulated the planning of the sub-total resection and radiotherapy preoperatively. Compared with control group, the amount of blood transfusion reduced significantly (P<0.05) and postoperative complications decreased. Conclusion MSCTA can imitate the surgical operations in multi-angle and supply the vital information for choosing the proper surgical approach, thereby reducing surgical trauma and postoperative complications.