Comparing the precision of the pSOFA and SIRS scores in predicting sepsis-related deaths among hospitalized children: a multi-center retrospective cohort study.

BACKGROUND: The latest sepsis definition includes both infection and organ failure, as evidenced by the sequential organ failure assessment (SOFA) score. However, the applicability of the pediatric SOFA score (pSOFA) is not yet determined. This study evaluated the effectiveness of both pSOFA and system inflammatory reaction syndrome (SIRS) scores in predicting sepsis-related pediatric deaths. METHODS: This is a retrospective multi-center cohort study including hospitalized patients <18 years old with diagnosed or not-yet-diagnosed infections. Multivariate analyses were carried out to evaluate risk factors for in-hospital mortality. According to Youden index (YI), three sub-categories of pSOFA were screened out and a new simplified pSOFA score (spSOFA) was formed. The effectiveness and accuracy of prediction of pSOFA, SIRS and spSOFA was retrieved from the area under the receiver operating characteristic curve (AUROC) and Delong's test. RESULTS: A total of 1,092 participants were eligible for this study, and carried a 23.4% in-hospital mortality rate. The 24-h elevated pSOFA score (24 h-pSOFA), bloodstream infection, and mechanical ventilation (MV) requirement were major risk factors associated with sepsis-related deaths. The AUROC analysis confirmed that the spSOFA provided good predictive capability in sepsis-related pediatric deaths, relative to the 24 h-pSOFA and SIRS. CONCLUSIONS: The pSOFA score performed better than SIRS in diagnosing infected children with high mortality risk. However, it is both costly and cumbersome. We, therefore, proposed spSOFA to accurately predict patient outcome, without the disadvantages. Nevertheless, additional investigations, involving a large sample population, are warranted to confirm the conclusion of this study.

Study on the Correlation between Urinary Sodium and Potassium Excretion and Blood Pressure in Adult Hypertensive Inpatients of Different Sexes.

Objective: This study aims to understand the difference in the influence of urinary sodium and potassium excretion on blood pressure in patients of different sexes with hypertension by analyzing the relationship between urinary sodium and potassium excretion and blood pressure. Methods: In this cross-sectional study, 606 hospitalized patients with essential hypertension were recruited from 16 hospitals in the Shanxi Province between June 2018 and December 2019. These patients were grouped by sex, with 368 males and 238 females. Basic information and relevant serum biochemical indexes of patients in the two groups were recorded. The 24-hour urinary sodium and potassium excretion were measured, and 24-hour ambulatory blood pressure monitoring was performed simultaneously. This was done to analyze and compare the relationship between urinary sodium and urinary potassium excretion and blood pressure in adult hospitalized patients of different sexes with hypertension. Results: The 24-hour urinary sodium excretion in male patients with hypertension was significantly higher than that in female patients (P < 0.001). There was no significant difference in 24-hour urinary potassium excretion between male patients with hypertension and female patients. Spearman correlation analysis showed that 24-hour urinary sodium excretion was positively correlated with 24-hour SBP and nSBP in male patients (P < 0.05), while 24-hour urinary potassium excretion was negatively correlated with 24-hour SBP and nSBP in male patients (P < 0.05). The 24-hour urinary sodium in female patients was significantly positively correlated with 24-hour SBP, 24-hour DBP, SBP, dDBP, nSBP, and nDBP (P < 0.01). The 24-hour urinary potassium was significantly negatively correlated with nSBP (P < 0.05). Multiple stepwise linear regression showed that 24-hour urinary sodium excretion was still significantly positively correlated with 24-hour SBP and nSBP in male patients with hypertension after adjusting for various confounding factors. Conclusion: High urinary sodium and low urinary potassium excretion are closely related to elevated blood pressure in adult patients with hypertension, and there are sex differences.

HIV-1-Specific CD11c+ CD8+ T Cells Display Low PD-1 Expression and Strong Anti-HIV-1 Activity.

Exhaustion of HIV-1-specific CD8+ T cells prevents optimal control of HIV-1 infection. Identifying unconventional CD8+ T cell subsets to effectively control HIV-1 replication is vital. In this study, the role of CD11c+ CD8+ T cells during HIV-1 infection was evaluated. The frequencies of CD11c+ CD8+ T cells significantly increased and were negatively correlated with viral load in HIV-1-infected treatment-naïve patients. HIV-1-specific cells were enriched more in CD11c+ CD8+ T cells than in CD11c- CD8+ T cells, which could be induced by HIV-1-derived overlapping peptides, marking an HIV-1-specific CD8+ T cell population. This subset expressed higher levels of activating markers (CD38 and HLA-DR), cytotoxic markers (granzyme B, perforin, and CD107a), and cytokines (IL-2 and TNF-α), with lower levels of PD-1 compared to the CD11c- CD8+ T cell subset. In vitro analysis verified that CD11c+ CD8+ T cells displayed a stronger HIV-1-specific killing capacity than the CD11c- counterparts. These findings indicate that CD11c+ CD8+ T cells have potent immunotherapeutic efficacy in controlling HIV-1 infection.

Single-Cell Transcriptomic Profiling of MAIT Cells in Patients With COVID-19.

Background: Mucosal-associated invariant T (MAIT) cells are considered to participate of the host immune response against acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection; however, single-cell transcriptomic profiling of MAIT cells in patients with COVID-19 remains unexplored. Methods: We performed single-cell RNA sequencing analyses on peripheral MAIT cells from 13 patients with COVID-19 and 5 healthy donors. The transcriptional profiles of MAIT cells, together with assembled T-cell receptor sequences, were analyzed. Flow cytometry analysis was also performed to investigate the properties of MAIT cells. Results: We identified that differentially expressed genes (DEGs) of MAIT cells were involved in myeloid leukocyte activation and lymphocyte activation in patients with COVID-19. In addition, in MAIT cells from severe cases, more DEGs were enriched in adaptive cellular and humoral immune responses compared with those in moderate cases. Further analysis indicated that the increase of cell cytotoxicity (killing), chemotaxis, and apoptosis levels in MAIT cells were consistent with disease severity and displayed the highest levels in patients with severe disease. Interestingly, flow cytometry analysis showed that the frequencies of pyroptotic MAIT cells, but not the frequencies of apoptotic MAIT cells, were increased significantly in patients with COVID-19, suggesting pyroptosis is one of leading causes of MAIT cell deaths during SARS-CoV-2 infection. Importantly, there were more clonal expansions of MAIT cells in severe cases than in moderate cases. Conclusions: The results of the present study suggest that MAIT cells are likely to be involved in the host immune response against SARS-CoV-2 infection. Simultaneously, the transcriptomic data from MAIT cells provides a deeper understanding of the immune pathogenesis of the disease.

The Correlation Between Urinary Sodium Excretion and Blood Pressure in Hospitalized Adult Patients with Hypertension.

INTRODUCTION: This study was designed to understand the baseline salt intake of adult patients with hypertension in Shanxi Province, and to analyze the correlation between urinary sodium excretion and blood pressure. METHODS: From June 2018 to December 2019, 16 hospitals with regional representativeness and experimental conditions in Shanxi Province were selected, and 643 eligible adult inpatients with primary hypertension were enrolled from these hospitals. The ages of patients ranged from 18 to 80 years. A 24-h ambulatory blood pressure monitoring was performed, and morning urine sodium concentration and 24-h urine sodium excretion were measured. The correlation between urinary sodium excretion and blood pressure in adult patients with hypertension was analyzed. RESULTS: The baseline salt intake of the adult patient participants with hypertension in Shanxi Province was 11.51 g/day. The average 24-h urinary sodium excretion of all observed subjects was 191.90 ± 98.18 mmol. The 24-h urinary sodium excretion and morning urinary sodium concentration were significantly positively correlated with systolic and diastolic blood pressure following adjustment of confounding factors, including gender, age, body weight, and smoking. CONCLUSION: The morning urine sodium concentration and 24-h urine sodium excretion were significantly positively correlated with blood pressure. High sodium excretion may be a risk factor for rhythm abnormalities in non-dipper pattern blood pressure. The control of urinary sodium concentration can thus be an important strategy for regulating abnormal blood pressure rhythm.

Role of non-inflammatory factors in intestinal fibrosis.

Intestinal fibrosis is a common complication of inflammatory bowel disease (IBD), resulting in strictures and ultimately obstruction, which is a significant clinical problem. Fibrosis is mainly triggered by local chronic inflammation and occurs when excessive extracellular matrix deposition is caused by activated mesenchymal cells. Despite the advance of anti-inflammatory therapies in IBD, the incidence and preventive strategies of intestinal fibrosis and strictures in IBD have not significantly changed over time. This shows that inflammation is necessary for fibrosis, but it does not necessarily affect the fibrotic progression. This review summarizes current knowledge about the non-inflammatory mechanisms implicated in the gut fibrotic process of IBD, which may pave the way for new mechanisms and anti-fibrotic therapies.

Chronic ethanol treatment of human hepatocytes inhibits the activation of the insulin signaling pathway by increasing cytosolic free calcium levels.

The present study aimed to investigate the effects of ethanol treatment on the induction of intracellular calcium ([Ca(2+)](i)) levels and the inhibition of the activation of the insulin signaling pathway in human hepatocytes. L­02 cells were treated with various concentrations of ethanol for different periods of time. Cell viability and alanine aminotransferase (ALT)/aspartate aminotransferase (AST) leakage in the culture supernatant were evaluated. Changes in [Ca(2+)](i) levels were detected by flow cytometry and confocal microscopy. Total RNA and protein were extracted to examine the mRNA and protein levels of insulin receptor substrate (IRS)1, IRS2, phosphatidylinositol 3­kinase (PI3K) and glucose transporter 2 (GLUT2) by reverse transcription-quantitative polymerase chain reaction (RT­qPCR) and western blot analysis, respectively. Furthermore, insulin was added to the ethanol­treated L­02 cells, and the phosphorylation levels of PI3K and protein kinase B (PKB) were determined by western blot analysis before and after Ca(2+) blockage. No significant changes were observed in cell viability, [Ca(2+)](i) levels and in the expression and phosphorylation levels of insulin signal transduction molecules when the L­02 cells were treated with 0.5 or 1% ethanol. However, treatment with 2 or 4% ethanol resulted in a significant decrease in cell viability and in the mRNA levels of IRS1, IRS2, PI3K (p85α) and GLUT2, as well as in an increase in ALT/AST leakage and in the [Ca(2+)](i) levels (P<0.05). The expression and phosphorylation levels of PI3K (p85α) and PKB were also inhibited by treatment with 2 or 4% ethanol. These cytological effects induced by ethanol treatment were partially reversed by Ca(2+) blockage. These results suggest that ethanol treatment inhibits the activation of the insulin signal transduction pathway in a dose­, time­ and Ca(2+)­dependent manner. The inhibition of IRS1/2, PI3K (p85α), PKB and GLUT2 expression and of PI3K (p85α) and PKB phosphorylation by the high concentrations of ethanol may be the core molecular mechanism of ethanol-induced insulin resistance, and may be related to the induction of [Ca(2+)](i) levels.

CagA(+) H. pylori induces Akt1 phosphorylation and inhibits transcription of p21(WAF1/CIP1) and p27(KIP1) via PI3K/Akt1 pathway.

OBJECTIVE: Cytotoxin-associated protein (CagA) of H. pylori has been confirmed to be closely associated with gastric inflammation and tumorigenesis, but the mechanism behind it is little understood. In this study, we try to determine roles of CagA(+) strain in activating PI3K/Akt1 signaling pathway, and affecting expression of p21(WAF1/CIP1) and p27(KIP1), and also in releasing IL-8 in host cells. METHODS: Akt1 phosphorylation and IL-8 levels of CagA(+) and CagA⁻ strain infected AGS cells were detected by ELISAs. Two quantitative RT-PCRs were established to measure p21(WAF1/CIP1) and p27(KIP1) mRNA levels in the CagA(+) and CagA⁻ strain infected cells. LY294002, an inhibitor of PI3K/Akt pathway, was used to define effect of the pathway in IL-8 release. RESULTS: CagA(+) strain could induce an obvious elevation of Akt1 phosphorylation in the infected AGS cells while CagA? strain failed to do so. The CagA(+) H. pylori strain infected AGS cells showed significant drops both in p21(WAF1/CIP1) and p27(KIP1) mRNA levels, whereas the CagA⁻ H. pylori strain caused a remarkable increase in p21(WAF1/CIP1) mRNA without affecting p27(KIP1) gene transcription in the AGS cells. Both the CagA(+) and CagA⁻ H. pylori strains enabled AGS cells to produce close elevated levels of IL-8, and the LY294002 block resulted in unexpected elevations of IL-8 levels. CONCLUSIONS: CagA can activate PI3K/Akt1 pathway that plays an inhibitory role in IL-8 release in H. pylori infected AGS cells. Activation of PI3K/Akt1 pathway and subsequent negative regulation of p21(WAF1/CIP1) and p27(KIP1) expression might be involved in CagA-associated carcinogenesis.

[Correlation between predominant pI genotypes, G120/A121 mutations and drug resistance].

OBJECTIVE: To analyze the predominant genotypes of outer membrane porin I (PI) and to determine the correlation between G120 as well as A121 mutations in PI proteins and drug resistance in Neisseria gonorrhoeae isolates in the local area. METHODS: A double PCR to simultaneously detect both pIA and pIB genes, was established in this study. The target amplification products were T-A cloned and then sequenced to determine the mutations at G120, A121 and the specificity of double PCR. By using acidity slip method and double agar dilution method, the beta-lactamase production and resistance to six antibiotics of pIA(+) and pIB(+) gonococcal isolates were detected. RESULTS: Double PCR could be used to accurately genotyping pI genes in all the tested gonococcal isolates with the sensitivity of 1 ng DNA template. In the 116 N. gonorrhoeae isolates, 30.2% (35/116) were pIA(+) strains and 69.8% (81/136) were pIB(+) strains. All the pIA(+) strains presented G120D/A121G double mutations (88.6%)or A121G single mutation (11.4%). 98.8% of the pIB(+) strains presented G120K/A121D (65.0%), G120K/A121G or G120N/A121D (13.8%) double mutations, and G120D/N/K single mutation (21.3%). 34.5% (40/116) of the isolates produced beta-lactamase, and the enzyme-produced rate (20%) in pIA(+) strains was significantly lower than that in pIB(+) strains (40.7%) with P < 0.05. No spectinomycin-resistant strains were identified but three ceftriaxone-resistant strains were presented. However, the resistance ratios to penicilin, tetramycin, ciprofloxacin and azithromycin of all the isolates were as high as 75.0% - 90.5%. 100% and 71.4% of the pIA(+) strains without beta-lactamase production and with G120 and/or A121 mutations were sensitive to penicillin and tetramycin, respectively. On the contrast, 100% of the pIB(+) strains without beta-lactamase production and with G120 and/or A121 mutations were resistant to both the two antibiotics. CONCLUSION: The established double PCR method could be used for fast and accurate genotyping of N. gonorrhoeae pI genes. The N. gonorrhoeae strains prevalent in the local areas mainly possessed pIB gene. Both spectinomycin and ceftriaxone could still be chosen to treat gonorrhea. The resistance enhancement caused by G120 and/or A121 mutations to penicillin and tetramycin was only presented in pIB(+) gonococci.

Involvement of Bombyx mori nucleopolyhedrovirus ORF41 (Bm41) in BV production and ODV envelopment.

Bombyx mori nucleopolyhedrovirus (BmNPV) ORF41 (Bm41), homologous to Ac52, is a gene present in most lepidopteran nucleopolyhedroviruses. Bm41 transcripts and encoded protein in BmNPV-infected cells can be detected from 3 and 6 h post-infection, respectively. Immunoassays have shown that Bm41 is not a viral structural protein and is detected in both the nuclei and cytoplasm of infected cells. A Bm41-disrupted virus (vBm(De)) and a repaired virus (vBm(Re)) were generated to investigate the function of Bm41. The results showed that Bm41 was essential for viral replication, and the disruption of Bm41 resulted in a much lower viral titer. Transmission electron microscopy revealed that disruption of Bm41 affected normal nucleocapsid envelopment and polyhedra formation in the nucleus. The disruption of Bm41 might severely affect odv-ec27 and polyhedrin expression. The disrupted virus reduced BmNPV infectivity in an LD(50) bioassay and took 18-23 h longer to kill larvae than wild-type virus in an LT(50) bioassay.

Vibrio vulnificus cytolysin induces apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells via caspase-9/3-dependent pathway.

Vibrio vulnificus cytolysin (VVC) is known to be a pore-forming toxin which shows cytotoxicity for mammalian cells in culture and induces apoptosis in endothelial cells. In order to determine whether VVC induces apoptosis in vascular endothelial cells and tumor cells, the cytotoxicity induced by recombinant VVC (rVVC) and its potential mechanism in HUVEC, SGC-7901 and SMMC-7721 cells were investigated. Our study demonstrated that rVVC induced the release of intracellular K(+) from all the target cells, yet lactate dehydrogenase was not released by rVVC. It indicates that osmotic lysis might not contribute to the cytolysin-induced cytotoxicity. The study also demonstrated that rVVC induced apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells in time- and dosage-dependent manners, which was associated with the activation of caspase-9 and -3, but not caspase-8. During the apoptotic process of the target cells, rVVC labeled with FITC was monitored to attach initially to the surface of the cells and entered the cytoplasma subsequently. These findings suggest that VVC may be not only a pore-forming toxin, but also a transmembrane toxin with powerful ability to induce apoptosis in human vascular endothelial cells and tumor cells.

Characterization of an early gene orf122 from Bombyx mori nucleopolyhedrovirus.

The open reading frame 122 of Bombyx mori nucleopolyhedrovirus (BmNPV) (Bm122) has been observed to be a conserved gene in the lepidopteran baculoviruses that have been completely sequenced so far. Its transcript was detected at 3 h post infection (h p.i.) and remained detectable at up to 96 h p.i. Temporal transcription analysis indicated that Bm122 is transcribed by host RNA polymerase. The size of the translational product of the Bm122 gene in Tn5B-1-4 cells was approximately 23 kDa, which is in agreement with the predicted value of 22.9 kDa, suggesting that no major posttranslational modification occurred in the primary protein product. The subcellular localization of Bm122 was studied using EGFP-Bm122, which revealed that Bm122 protein was accumulated within the nuclear region of virus-infected BmN cells. All these results suggest that Bm122 is an early gene encoding a protein that functions in the nucleus.

Characterization of a Bombyx mori nucleopolyhedrovirus with Bmvp80 disruption.

A BmNPV Bacmid with the Bmvp80 gene disrupted was constructed using the ET-recombination system in Escherichia coli to investigate the role of Bmvp80 during the baculovirus life cycle. Disruption of Bmvp80 resulted in single cell infection phenotype, whereas a rescue BmBacmid restored budded virus titers to wild type levels; however, the homologous gene Ac104 (Acvp80) from AcMNPV could not complement the BmBacmid lacking a functional Bmvp80 gene. Electron microscopy of cells transfected with BmNPV lacking functional Bmvp80 revealed that the number of nucleocapsids was markedly lower. These results suggest that Bmvp80 is essential for normal budded virus production and nucleocapsid maturation, and is functionally divergent between baculovirus species.

Bombyx mori nucleopolyhedrovirus ORF56 encodes an occlusion-derived virus protein and is not essential for budded virus production.

Bombyx mori nucleopolyhedrovirus ORF56 (Bm56) is a baculovirus core gene that is highly conserved in all baculoviruses that have had their genomes sequenced to date. Its transcripts in BmNPV-infected cells could be detected from 12 h post-infection (p.i.) and the encoded protein could be detected at 16 h p.i. by using a polyclonal antibody against glutathione S-transferase-Bm56 fusion protein. Western blot analysis showed that Bm56 is a structural component of the occlusion-derived virus nucleocapsid. Subsequent confocal microscopy revealed that Bm56 was distributed in the outer nuclear membrane and the intranuclear region of infected cells. To investigate the role of Bm56 in virus replication, a Bm56-knockout bacmid of BmNPV was constructed via homologous recombination in Escherichia coli. The Bm56 deletion had no effect on budded virus (BV) production in cultured cells; however, the deletion affected occlusion-body morphogenesis. A larval bioassay demonstrated that the Bm56 deletion did not reduce infectivity, whereas it resulted in a 50 % lethal time that was 16-18 h longer than that of the wild-type bacmid at every dose used in this study. These results indicate that Bm56 facilitates efficient virus production in vivo; however, it is not essential for BV production in vitro.