Azvudine and nirmatrelvir-ritonavir in hospitalized patients with moderate-to-severe COVID-19: Emulation of a randomized target trial.

To examine the effectiveness of azvudine and nirmatrelvir-ritonavir in treating hospitalized patients with moderate-to-severe COVID-19. We emulated a target trial with a multicenter retrospective cohort of hospitalized adults with moderate-to-severe COVID-19 without contraindications for azvudine or nirmatrelvir-ritonavir between December 01, 2022 and January 19, 2023 (during the Omicron BA.5.2 variant wave). Exposures included treatment with azvudine or nirmatrelvir-ritonavir for 5 days versus no antiviral treatment during hospitalization. Primary composite outcome (all-cause death and initiation of invasive mechanical ventilation), and their separate events were evaluated. Of the 1154 patients, 27.2% were severe cases. In the intent-to-treat analyses, azvudine reduced all-cause death (Hazard ratio [HR]: 0.31; 95% CI: 0.12-0.78), and its composite with invasive mechanical ventilation (HR: 0.47; 95% CI: 0.24-0.92). Nirmatrelvir-ritonavir reduced invasive mechanical ventilation (HR: 0.42; 95% CI: 0.17-1.05), and its composite with all-cause death (HR: 0.38; 95% CI: 0.18-0.81). The study did not identify credible subgroup effects. The per-protocol analyses and all sensitivity analyses confirmed the robustness of the findings. Both azvudine and nirmatrelvir-ritonavir improved the prognosis of hospitalized adults with moderate-to-severe COVID-19.

Not Only High Number and Specific Comorbidities but Also Age Are Closely Related to Progression and Poor Prognosis in Patients With COVID-19.

Background: Some patients with comorbidities and rapid disease progression have a poor prognosis. Aim: We aimed to investigate the characteristics of comorbidities and their relationship with disease progression and outcomes of COVID-19 patients. Methods: A total of 718 COVID-19 patients were divided into five clinical type groups and eight age-interval groups. The characteristics of comorbidities were compared between the different clinical type groups and between the different age-interval groups, and their relationships with disease progression and outcomes of COVID-19 patients were assessed. Results: Approximately 91.23% (655/718) of COVID-19 patients were younger than 60 years old. Approximately 64.76% (465/718) had one or more comorbidities, and common comorbidities included non-alcoholic fatty liver disease (NAFLD), hyperlipidaemia, hypertension, diabetes mellitus (DM), chronic hepatitis B (CHB), hyperuricaemia, and gout. COVID-19 patients with comorbidities were older, especially those with chronic obstructive pulmonary disease (COPD) and cardiovascular disease (CVD). Hypertension, DM, COPD, chronic kidney disease (CKD) and CVD were mainly found in severe COVID-19 patients. According to spearman correlation analysis the number of comorbidities was correlated positively with disease severity, the number of comorbidities and NAFLD were correlated positively with virus negative conversion time, hypertension, CKD and CVD were primarily associated with those who died, and the above-mentioned correlation existed independently of age. Risk factors included age, the number of comorbidities and hyperlipidaemia for disease severity, the number of comorbidities, hyperlipidaemia, NAFLD and COPD for the virus negative conversion time, and the number of comorbidities and CKD for prognosis. Number of comorbidities and age played a predictive role in disease progression and outcomes. Conclusion: Not only high number and specific comorbidities but also age are closely related to progression and poor prognosis in patients with COVID-19. These findings provide a reference for clinicians to focus on not only the number and specific comorbidities but also age in COVID-19 patients to predict disease progression and prognosis. Clinical Trial Registry: Chinese Clinical Trial Register ChiCTR2000034563.

Lymphocyte subsets with the lowest decline at baseline and the slow lowest rise during recovery in COVID-19 critical illness patients with diabetes mellitus.

BACKGROUND: Host dysregulation of immune response was highly involved in the pathological process of Coronavirus disease 2019 (COVID-19), especially COVID-19 severe cases with DM. AIM: In this study we aimed at the dynamic change of peripheral lymphocyte and subsets during COVID-19 covery. METHODS: The peripheral lymphocyte and subsets of 95 confirmed cases with COVID-19 from baseline to four weeks were compared between critical illness and non-critical illness cases with or without DM. RESULTS: The dynamic characteristics of lymphocyte and subsets in COVID-19 patients was that it reduced significantly at one week, rapidly elevated to the peak at two weeks after onset, then gradually declined during recovery. The COVID-19 critical illness patients with DM had the lowest decline at one week and the slow lowest rise at two weeks after onset, while COVID-19 non-critical illness patients with DM had the rapid highest rise at two weeks after onset, both of them had similar lymphocyte and subsets at five weeks after onset and lower than those patients without DM. CONCLUSIONS: These findings provide a reference for clinicians that for COVID-19 patients with DM and the lowest decline of lymphocyte and subsets, immunomodulatory therapy as soon as possible might avoid or slow down disease progression; moreover for COVID-19 critical illness patients with or without DM and non-critical illness patients with DM, continuous immunomodulatory therapy in later stages of disease might speed up virus clearance, shorten hospital stay, improve disease prognosis in COVID-19 critical illness patients with DM.

Alternative complement pathway is activated in the brains of scrapie-infected rodents.

Activation of complement system in central nervous system (CNS) of the patients suffering from prion diseases or animal models infected with prion agents experimentally is reported repeatedly, but which pathways are involved in the complement system during prion infection is not well documented. Here, we evaluated the level of complement factor B (CFB), which is the key factor that triggers alterative pathway (AP) of complement in the brain tissues of scrapie-infected mice with various methodologies. We found that the levels of mRNA and protein of CFB significantly increased in the brain tissues of scrapie-infected mice. Morphologically, the increased CFB-specific signal overlapped with the elevated C3 signal in brain sections of scrapie-infected mice, meanwhile overlapped with damaged neurons and activated microglia, but not with the proliferative astrocytes. Additionally, the level of complement factor P (CFP), the key positive regulator of AP, also increased remarkably in the brain tissues of infected mice. The transcriptional levels of CD55 and CD46, two negative regulators of AP, decreased without significance in brain tissues of scrapie-infected mice at the terminal stage. However, the mRNA and protein levels of CFH, another negative regulator of AP, increased. Through the dynamic analyses of the expressions of CFB, CFP, and CFH in brain sections of 139A-infected mice, which were collected at different time-points during incubation period, illustrated time-dependent increase levels of each factor during the incubation period of scrapie infection. Taken together, our data here demonstrate that the AP of complement cascade is activated in the CNS microenvironment during prion infection.

Aberrant alterations of the expressions and S-nitrosylation of calmodulin and the downstream factors in the brains of the rodents during scrapie infection.

The aberrant alterations of calmodulin (CaM) and its downstream substrates have been reported in some neurodegenerative diseases, but rarely described in prion disease. In this study, the potential changes of Ca2+/CaM and its associated agents in the brains of scrapie agent 263K-infected hamsters and the prion infected cell line SMB-S15 were evaluated by various methodologies. We found that the level of CaM in the brains of 263K-infected hamsters started to increase at early stage and maintained at high level till terminal stage. The increased CaM mainly accumulated in the regions of cortex, thalamus and cerebellum of 263K-infected hamsters and well localization of CaM with NeuN positive cells. However, the related kinases such as total and phosphorylated forms of CaMKII and CaMKIV, as well as the downstream proteins such as CREB and BDNF in the brain of 263K-infected hamsters were decreased. Further analysis showed a remarkable increase of S-nitrosylated (SNO) form of CaM in the brains of 263K-infected hamsters. Dynamic analysis of S-nitrosylated CaM showed the SNO form of CaM abnormally increases in a time-dependent manner during prion infection. Compared with that of the normal partner cell line SMB-PS, the CaM level in SMB-S15 cells was increased, meanwhile, the downstream proteins, such as CaMKII, p-CaMKII, CREB, as well as BDNF, were also increased, especially in the nucleic fraction. No SNO-CaM was detected in the cell lines SMB-S15 and SMB-PS. Our data indicate an aberrant increase of CaM during prion infection in vivo and in vitro.

Remarkable increases of α1-antichymotrypsin in brain tissues of rodents during prion infection.

α1-Antichymotrypsin (α1-ACT) belongs to a kind of acute-phase inflammatory protein. Recently, such protein has been proved exist in the amyloid deposits which is the hallmark of Alzheimer's disease, but limitedly reported in prion disease. To estimate the change of α1-ACT during prion infection, the levels of α1-ACT in the brain tissues of scrapie agents 263K-, 139A- and ME7-infected rodents were analyzed, respectively. Results shown that α1-ACT levels were significantly increased in the brain tissues of the three kinds of scrapie-infected rodents, displaying a time-dependent manner during prion infection. Immunohistochemistry assays revealed the increased α1-ACT mainly accumulated in some cerebral regions of rodents infected with prion, such as cortex, thalamus and cerebellum. Immunofluorescent assays illustrated ubiquitously localization of α1-ACT with GFAP positive astrocytes, Iba1-positive microglia and NeuN-positive neurons. Moreover, double-stained immunofluorescent assays and immunohistochemistry assays using series of brain slices demonstrated close morphological colocalization of α1-ACT signals with that of PrP and PrPSc in the brain slices of 263K-infected hamster. However, co-immunoprecipitation does not identify any detectable molecular interaction between the endogenous α1-ACT and PrP either in the brain homogenates of 263K-infected hamsters or in the lysates of prion-infected cultured cells. Our data here imply that brain α1-ACT is increased abnormally in various scrapie-infected rodent models. Direct molecular interaction between α1-ACT and PrP seems not to be essential for the morphological colocalization of those two proteins in the brain tissues of prion infection.

[Analysis of piperidinium ionic liquid cations by ion chromatography with direct conductivity detection].

A method of ion chromatography with direct conductivity detection was established to determine three piperidinium ionic liquid cations, i. e. N-methyl, ethyl piperidinium cation ([MEPi]+), N-methyl, propyl piperidinium cation ([MPPi]+) and N-methyl, butyl piperidinium cation ([MBPi]+). The effects of eluent types, eluent concentration and column temperature on the retention of the cations were investigated with sulfonic acid base cation exchange column using ethylenediamine-citric acid-acetonitrile as eluent. The results indicated that, with the increase of column temperature, the retention times of piperidinium cations were reduced, so the retention process of piperidinium cations was exothermic. The retentions of piperidinium cation homologue accorded with carbon number rule. The successful separation of the three piperidinium cations within 7 min was achieved using the optimized eluent of 0.2 mmol/L ethylenediamine-0.3 mmol/L citric acid-3% acetonitrile (pH 4.4) at a flow rate of 1.0 mL/min and the column temperature of 30 degrees C. Under these conditions, the detection limits at a signal-to-noise ratio of 3 for [MEPi]+, [MPPi] and [MBPi]+ were 0.14, 0.20 and 0.56 mg/L, respec-tively. The relative standard deviations (n = 5) for peak areas were less than 1.2%. The method has been applied to the determination of piperidinium ionic liquids synthesized in chemical laboratory with the spiked recoveries of 97.6% to 105.1%. The method is accurate, reliable, rapid, and has better practicability.

[Determination of ionic liquid anions of trifluoroacetate, thiocyanate, tetrafluoroborate and trifluoromethanesulfonate by ion-pair chromatography with direct conductivity detection].

An ion-pair chromatographic method with direct conductivity detection was established on a reversed-phase silica-based column for the simultaneous determination of ionic liquid anions of trifluoroacetate, thiocyanate, tetrafluoroborate and trifluoromethanesulfonate. The analytes were separated using a mobile phase of ion pair reagent-malic acid-acetonitrile on the Diamonsil C18 column. The effects of mobile phase composition and column temperature on the retention of the anions were investigated. The optimized chromatographic conditions were as follows: 0.15 mmol/L tetrabutylammonium hydroxide-0.099 mmol/L malic acid-20% (v/v) acetonitrile aqueous solution (pH 6.5) as mobile phase, a column temperature of 25 degrees C. Under the optimal conditions, the baseline separation of trifluoroacetate, thiocyanate, tetrafluoroborate and trifluoromethanesulfonate was achieved without any interference by other ordinary anions (fluoride, chloride, bromide, nitrate, sulfate). The detection limits (S/N = 3) were 0.21, 0.07, 0.36 and 0.12 mg/L for trifluoroacetate, thiocyanate, tetrafluoroborate and trifluoromethanesulfonate, respectively. The method has been applied to the determination of the four anions in ionic liquids. The spiked recoveries of the anions were from 95.0% to 104.6%. The results demonstrate that the convenience, rapidity, sensitivity and accuracy are fit for the requirements of quantitative analysis of trifluoroacetate, thiocyanate, tetrafluoroborate and trifluoromethanesulfonate in ionic liquids.

Serum protein profiling of smear-positive and smear-negative pulmonary tuberculosis using SELDI-TOF mass spectrometry.

The focus of this study was to detect novel sera biomarkers for smear-positive and smear-negative pulmonary tuberculosis and to establish respective diagnostic models using the surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) technique. A total of 155 sera samples from smear-positive pulmonary tuberculosis (SPPTB) and smear-negative pulmonary tuberculosis (SNPTB) patients and non-tuberculosis (non-TB) controls were analyzed with SELDI-TOF MS. The study was divided into a preliminary training set and a blinded testing set. A classification tree of spectra derived from 31 SPPTB patients, 22 SNPTB patients, and 42 non-TB controls were used to develop an optimal classification tree that discriminated them respectively in the training set. Then, the validity of the classification tree was challenged with another independent blinded testing set, which included 20 SPPTB patients, 14 SNPTB patients, and 26 non-TB controls. SNPTB patients and non-TB controls also were analyzed alone using the same method. The optimal decision tree model with a panel of nine biomarkers with mass:charge ratios (m/z) of 4821.45, 3443.22, 9284.93, 4473.86, 4702.84, 3443.22, 5343.26, 3398.27, and 3193.61 determined in the training set could detect 93.55%, 95.46%, and 88.09% accuracy for classifying SPPTB patients, SNPTB patients, and non-TB controls specimens, respectively. Validation of an independent, blinded testing set gave an accuracy of 80.77% for controls, 75.00% for SPPTB, and 71.43% for SNPTB samples using the same classification tree. With the peaks displaying differences between SNPTB patients and non-TB controls, a simplified dendrogram (m/z 4821.45, 4792.74) demonstrated classification efficacy of 85.94% (sensitivity 86.36% and specificity 85.71%) for distinguishing SNPTB patients from non-TB controls. The independent blinded testing set containing 14 SNPTB patients and 26 non-TB controls gained an accuracy of 81.59% (sensitivity 78.57% and specificity 84.62%) for diagnosing SNPTB. Special proteins/peptides may change in SPPTB and SNPTB patients and those changes may be used to distinguish them with the proper discriminant analytical method and to pursue and identify some involved proteins underlying the biological process of tuberculosis.