Background Effective removal of pathogenic bacteria is key to improving the prognosis of sepsis. Polymorphonuclear neutrophils (PMNs) are the most important components of innate cellular immunity and play vital roles in clearing pathogenic bacteria. However, the metabolic characteristics and immunomodulatory pathways of PMNs during sepsis have not been investigated. In the present study, we explored the immune metabolism characteristics of PMNs and the mechanism by which neutrophilic glycolysis is regulated during sepsis. Methods Metabolomics analysis was performed on PMNs isolated from 14 septic patients, 26 patients with acute appendicitis, and 19 healthy volunteers. Transcriptome analysis was performed on the PMNs isolated from the healthy volunteers and the patients with sepsis to assess glycolysis and investigate its mechanism. Lipopolysaccharide (LPS) was used to stimulate the neutrophils isolated from the healthy volunteers at different time intervals to build an LPS-tolerant model. Chemotaxis, phagocytosis, lactate production, oxygen consumption rate (OCR), and extracellular acidification rate (ECAR) were evaluated. Results Transcriptomics showed significant changes in glycolysis and the mTOR/HIF-1α signaling pathway during sepsis. Metabolomics revealed that the Warburg effect was significantly altered in the patients with sepsis. We discovered that glycolysis regulated PMNs’ chemotaxis and phagocytosis functions during sepsis. Lactate dehydrogenase A (LDHA) downregulation was a key factor in the inhibition of glycolysis in PMNs. This study confirmed that the PI3K/Akt-HIF-1α pathway was involved in the LDHA expression level and also influenced PMNs’ chemotaxis and phagocytosis functions. Conclusions The inhibition of glycolysis contributed to neutrophil immunosuppression during sepsis and might be controlled by PI3K/Akt-HIF-1α pathway-mediated LDHA downregulation. Our study provides a scientific theoretical basis for the management and treatment of patients with sepsis and promotes to identify therapeutic target for the improvement of immune function in sepsis.

Congenital ptosis has varying degrees of impact on the visual development or psychological health of a child depending on its severity. Some controversies and misconceptions remain regarding the management of congenital ptosis in children. Particularly, the accurate diagnosis of the severity of congenital ptosis in younger children, assessment of the visual developmental status of the child, optimal timing of surgery, and treatment choice are still issues in clinical practice that need to be explored. This report presents a comprehensive review of these aspects of the correction of congenital ptosis to provide a valuable reference for clinical practice. Our review shows that currently used surgical procedures for ptosis may result in over- or under-correction to varying degrees. The differences may be due to the physical condition and age of the child and the degree of cooperation during preoperative examination and assessment, resulting in inaccurate results. Alternatively, intraoperative swelling and bleeding may lead to errors in the values recorded by the surgeon. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Background: Since the outbreak of coronavirus disease (COVID-19) in December 2019 in Wuhan, it has spread rapidly worldwide. We aimed to establish and validate a nomogram that predicts the probability of coronavirus-associated acute respiratory distress syndrome (CARDS). Methods: In this single-centre, retrospective study, 261 patients with COVID-19 were recruited using positive reverse transcription??? polymerase chain reaction tests for severe acute respiratory syndrome coronavirus 2 in Tongji Hospital at Huazhong University of Science and Technology (Wuhan, China). These patients were randomly distributed into the training cohort (75%) and the validation cohort (25%). The factors included in the nomogram were determined using univariate and multivariate logistic regression analyses based on the training cohort. The area under the receiver operating characteristic curve (AUC), consistency index (C-index), calibration curve, and decision curve analysis (DCA) were used to evaluate the efficiency of the nomogram in the training and validation cohorts. Results: Independent predictive factors, including fasting plasma glucose, platelet, D-dimer, and cTnI, were determined using the nomogram. In the training cohort, the AUC and concordance index were 0.93. Similarly, in the validation cohort, the nomogram still showed great distinction (AUC: 0.92) and better calibration. The calibration plot also showed a high degree of agreement between the predicted and actual probabilities of CARDS. In addition, the DCA proved that the nomogram was clinically beneficial. Conclusion: Based on the results of laboratory tests, we established a predictive model for acute respiratory distress syndrome in patients with COVID-19. This model shows good performance and can be used clinically to identify CARDS early.

PURPOSE:Acute kidney injury (AKI) is a common organ dysfunction in ICU and up to now there is no good way to predict the AKI progression and patient prognosis. Blood electrolyte tests are common in ICU, but there are few studies on early blood electrolytes and the AKI progression and patient prognosis. Therefore, we concentrated on the serum sodium and potassium levels before AKI diagnosis and evaluated the relationship between serum sodium and potassium levels and the severity and prognosis of AKI. METHODS:This study included data of all patients from the MIMIC-III. We used the urine output criteria in the KDIGO as diagnostic criteria for oliguric AKI. Patients admitted to the ICU several times only included their initial ICU admission results. Patients younger than 18 years old, diagnosed with AKI stage 3, ICU stays less than 24 h or without corresponding laboratory results or data were excluded. The included patients were divided into four groups based on the interquartile range of serum sodium and potassium. We evaluated the serum sodium and potassium levels before AKI diagnosis and AKI severity and prognosis through retrospective analysis. RESULTS:Patients with serum potassium > 4.6 mmol/L were more likely to progress to AKI stage 3 or death than patients with serum potassium ≤ 4.6 mmol/L (overall p < 0.0001). Patients with sodium < 137 mmol/L or > 141 mmol/L had a higher risk of progressing to AKI stage 3 (overall p = 0.00023) and risk of death (overall p < 0.0001) than other patients. In the Cox regression model, after adjusting for age, sex, and BMI, serum sodium or potassium were associated with AKI progression and prognosis (p < 0.01). After continuing to adjust for comorbidities, serum potassium was still associated with AKI progression and prognosis (p < 0.01), but serum sodium was only associated with prognosis (p = 0.027). After adjusting for other indicators, there was no statistically significant correlation between serum sodium or potassium and AKI progression and prognosis. After adjusting for serum sodium or potassium, the corresponding results were not significantly different from those before adjustment. CONCLUSION:This study found that abnormal serum sodium or potassium levels before AKI diagnosis were more likely to lead to AKI progression and poor prognosis, of which lower serum sodium and higher serum potassium were more likely to progress to AKI stage 3 or death.

Adult-onset Still's disease (AOSD) is a systemic, multifactorial, autoinflammatory disease for which the etiopathogenesis is not well understood. Given the similarities in clinical and laboratory features between this disease and sepsis, and the differences in treatment strategies for these two diseases, specific diagnostic markers are crucial for the correct diagnosis and management of AOSD. Previous studies have shown plasma heparin-binding protein (HBP) is a promising potential biomarker for AOSD; thus, this study aimed to detect serum HBP levels in patients with AOSD or sepsis to assess its potential as a biomarker for differential diagnosis. We found that serum HBP levels were significantly higher in patients with active AOSD than that in those with inactive AOSD. Patients with sepsis had higher serum HBP levels compared with those who had active or inactive AOSD. We calculated the area under the receiver operating characteristic (ROC) curve to assess whether HBP could be used to differentiate active from inactive AOSD; this was 0.811 with sensitivity 0.650, specificity 0.811, and cutoff HBP value of 35.59 ng/ml. The area under the ROC curve for HBP as a biomarker to differentiate AOSD from sepsis was 0.653, with sensitivity 0.759, and specificity 0.552, and cutoff HBP value of 65.1 ng/ml. Taken together, the results of our study suggest that serum HBP could be a useful diagnostic biomarker to evaluate disease activity in patients with AOSD, and to differentiate AOSD from sepsis.

The study of cisplatin sensitivity is the key to the development of ovarian cancer treatment strategies. Mitochondria are one of the main targets of cisplatin, its self-clearing ability plays an important role in determining the fate of ovarian cancer cells. First, we proved that the sensitivity of ovarian cancer cells to cisplatin depends on mitophagy, and p62 acts as a broad autophagy receptor to regulate this process. However, p62′s regulation of mitophagy does not depend on its location on the mitochondria. Our research shows that the mutation of the UBA domain of p62 increases the localisation of HK2 on the mitochondria, thereby increasing the phosphorylated ubiquitin form of parkin, then stabilising the process of mitophagy and ultimately cell survival. Collectively, our results showed that a mutation in the UBA domain of p62 regulates the level of apoptosis stimulated by cisplatin in ovarian cancer.

OBJECTIVE:To explore special coagulation characteristics and anticoagulation management in extracorporeal membrane oxygenation (ECMO)-assisted patients with coronavirus disease 2019 (COVID-19). DESIGN:Single-center, retrospective observation of a series of patients. PARTICIPANTS:Laboratory-confirmed severe COVID-19 patients who received venovenous ECMO support from January 20-May 20, 2020. INTERVENTIONS:This study analyzed the anticoagulation management and monitoring strategies, bleeding complications, and thrombotic events during ECMO support. MEASUREMENTS AND MAIN RESULTS:Eight of 667 confirmed COVID-19 patients received venovenous ECMO and had an elevated D-dimer level before and during ECMO support. An ECMO circuit pack (oxygenator and tubing) was replaced a total of 13 times in all 8 patients, and coagulation-related complications included oxygenator thrombosis (7/8), tracheal hemorrhage (5/8), oronasal hemorrhage (3/8), thoracic hemorrhage (3/8), bleeding at puncture sites (4/8), and cannulation site hemorrhage (2/8). CONCLUSIONS:Hypercoagulability and secondary hyperfibrinolysis during ECMO support in COVID-19 patients are common and possibly increase the propensity for thrombotic events and failure of the oxygenator. Currently, there is not enough evidence to support a more aggressive anticoagulation strategy.

Purpose: A lack of early diagnostic biomarkers and therapeutic targets has led to poor prognosis for gastric cancer patients. However, the analysis of cancer-associated genomic data has been shown to be effective in identifying potential markers. Recently, the long noncoding RNA LINC00365 and SCGB2A1 gene (as known as mammaglobin B) were predicted to be co-expressed in gastric cancer based on the Gene Expression Omnibus database. However, their precise role in gastric cancer tumors is still not clear. Methods: The expressions of LINC00365 and SCGB2A1 in gastric cancer tissues were investigated using qPCR and their expressions were detected in a gastric cancer tissue microarray by in situ hybridization and immunohistochemical staining. The functions of LINC00365 in BGC-823 and MGC-803 gastric cancer cells were tested using the MTT assay, flow cytometry, colony formation assay, EDU staining, immunofluorescence and luciferase assay. Results: We found that LINC00365 and SCGB2A1 mRNA were both expressed at low levels in 30 cases of gastric cancer. Gastric cancer tissue microarray analysis indicated that LINC00365 and SCGB2A1 were expressed at low levels in tumor tissue, and low expression of both factors correlated with shorter survival time. Functional studies showed that LINC00365 overexpression significantly inhibited gastric cancer cell viability through the impairment of proliferation rather than the promotion of apoptosis. Furthermore, overexpressed LINC00365 upregulated SCGB2A1 in gastric cancer cell lines. Immuno-fluorescence and luciferase assay analysis indicated that LINC00365 overexpression inhibited the NF-kappa B prosurvival signaling pathway. Consistent with the effects of LINC00365, SCGB2A1 upregulation also reduced cell survival and inactivated NF-kappa B. Conclusion: Collectively, our findings revealed that SCGB2A1 may be the target coding protein regulated by LINC00365 in gastric cancer. LINC00365 and SCGB2A1 may function as tumor suppressors and may serve as potential prognostic and therapeutic markers in gastric cancer treatment.

The emergence of resistance to chemotherapy drugs in patients with ovarian cancer is still the main cause of low survival rates. The present study aimed to identify key genes that may provide treatment guidance to reduce the incidence of drug resistance in patients with ovarian cancer. Original data of chemotherapy sensitivity and chemoresistance of ovarian cancer were obtained from the Gene Expression Omnibus dataset GSE73935. Differentially expressed genes (DEGs) between sensitive and resistant ovarian cancer cell lines were screened by Empirical Bayes methods. Overlapping DEGs between four chemoresistant groups were identified by Venn map analysis. Protein-protein interaction networks were also constructed, and hub genes were identified. The hub genes were verified by in vitro experiments as well as The Cancer Genome Atlas data. Results from the present study identified eight important genes that may guide treatment decisions regarding chemotherapy regimens for ovarian cancer, including epidermal growth factor-like repeats and discoidin I-like domains 3, NRAS proto-oncogene, hyaluronan and proteoglycan link protein 1, activated protein C receptor, CD53, cyclin-dependent kinase inhibitor 2A, insulin-like growth factor 1 receptor and roundabout guidance receptor 2 genes. Their expressions were found to have an impact on the prognosis of different treatment groups (cisplatin, paclitaxel, cisplatin + paclitaxel, cisplatin + doxorubicin and cisplatin + topotecan). The results indicated that these genes may minimise the occurrence of ovarian cancer drug resistance and may provide effective treatment options for patients with ovarian cancer.

Cisplatin is a platinum-based first-line drug for treating ovarian cancer. However, chemotherapy tolerance has limited the efficacy of cisplatin for ovarian cancer patients. Research has demonstrated that cisplatin causes changes in cell survival and death signaling pathways through its interaction with macromolecules and organelles, which indicates that investigation into the DNA off-target effects of cisplatin may provide critical insights into the mechanisms underlying drug resistance. The multifunctional protein p62 works as a signaling hub in the regulation of pro-survival transcriptional factors NF-κB and Nrf2 and connects autophagy and apoptotic signals, which play important roles in maintaining cell homeostasis. In this review, we discuss the role of p62 in cisplatin resistance by exploring p62-associated signaling pathways based on current studies and our work. Insights into these resistance mechanisms may lead to more effective therapeutic strategies for ovarian cancer by targeting p62.

Objective: To resect benign tumors on the palpebral margin using an improved minimally invasive surgery, and to observe the outcome and analyze the possible healing mechanism of this improved surgical technique. Methods: Fifty-five patients with a benign tumor on the palpebral margin measuring 2- to 10 mm in diameter were included in this study. The tumors were resected along their edge and basal layer, causing minimal damage to the surrounding structures. Postoperative outcome measures included the following: the wound status, epithelial healing, eyelash growth, recurrence, and complications. Postoperative follow-up time points were 1 day, 7 days, 14 days, 1 month, 3 months, and 12 months. Results: The wound status ranged from edema to contraction in the early stage postoperatively. The eyelashes that were excised during the operation began to grow from 7 to 14 days postoperatively, and the epithelium began to close simultaneously. From 14 days to 3 months postoperatively, the eyelash and epithelium grew completely, and the scar disappear gradually. Except in 1 patient with trichiasis at 3 months and one patient with tumor recurrence at 12 months postoperatively, there were no other complications, such as ablepharon deformities, alopecia palpebralis, etc. Conclusion: Our improved minimally invasive technique not only resected the tumor effectively but also preserved the important anatomical structure of the palpebral margin. The favorable outcomes resulted from the mechanism of wound healing. This new surgical method is worth implementing in clinical practice.

Low binding affinity and lack of therapy functions limit tumor targeting peptide applications in the biomedical field. Herein, we successfully modified a previous phage display derived Glypican-3 (GPC3) binding peptide (GBP) on the surface of a Fe3O4 Core/Au shell nanocomplex (FANP) to improve GBP binding affinity and enhance FANP tumor photothermal therapy (PTT) efficacy. As a result, GBP-FANP showed improved avidity to GPC-3 (Apparent Kd = 396.3 ± 70.8 nM) compared to that of GPB (Apparent Kd = 735.2 ± 53.6 nM). After intravenous administration, GBP-FANP was found specifically accumulated in GPC-3 positive HepG2 tumors and peaked at 24 h post-injection as observed by magnetic resonance imaging (MRI)/photoacoustic (PA)/fluorescent imaging. Moreover, HepG2 tumors that received GBP-FANP treatment were significantly inhibited with laser irradiation (630 nm, 1 W cm-2, 10 min). In conclusion, our present strategy provides a way of improving peptide ligand avidity with nanotechnology for cancer theranostics applications.

Purpose: This study investigated the expression of HIF-1α and Robo4 in the retinas of streptozotocin (STZ)-induced mice and determined the expression correlation of these two factors in early diabetic retinas in vivo. Methods: A high-fat diet together with STZ stimulated type 2 diabetes mellitus (DM). HE staining was used to observe the morphologic features of the retinas following 4, or 8 weeks of hyperglycemia. Immunofluorescence was carried out to analyze the expression of HIF-1α and Robo4 in the retinas at different time points. HIF-1α and Robo4 mRNA and protein expressions were quantified by real-time PCR and western blot. Results: The arrangements of the retinal nerve fiber layer (NFL) and the ganglion cell layer (GCL) were slightly turbulent in the 4-week old diabetic mice, which became aggravated by NFL edema and cytoplasmic vacuoles in the 8-week old group. In the 4-week old group, HIF-1α was expressed slightly higher in NFL and GCL, and Robo4 expression increased in NFL and GCL. In the 8-week old diabetic retinas, HIF-1α expression was enhanced in NFL, GCL, and the outer plexiform layer (OPL); Robo4 expression increased apparently in NFL and GCL. HIF-1α and Robo4 mRNA and protein expressions were also increased slightly in the 1-week old retinas and significantly after 4 and 8 weeks. Conclusions: With aggravating retina structure turbulence in DM mice, both HIF-1α and Robo4 expressions were increased and mainly concentrated in the GCL, INL, and OPL, suggesting a regulatory role of HIF-1α on Robo4 and their combined effect on DM retina damage in vivo.

Objective: Massive obstetric hemorrhage (MOH) is one of the major causes of maternal morbidity and mortality. The purpose of our study was to investigate the risk factors of MOH and further explore the effect of MOH on prognosis. Study design: Retrospective study of maternal near miss (MNM) with MOH was managed in a maternity center during a 10-year study period. Risk factors including maternal age, gestational age, weekend delivery, polyembryony, and comorbidities were analyzed. Maternal outcomes including disseminated intravascular coagulopathy (DIC), acute renal failure, abdominal hematoma, need for transfusion of blood products, shock, MODS, stroke, cardiac arrest, maternal mortality, neonatal mortality, hysterectomy, and abortion were also analyzed. Other clinical outcomes included ICU stay and hospital stay. Results: A total of 283 women with MNM were included in this study. MOH was present in 15.5%, shock was present in 7.07%, and DIC was present in 2.47%. The rate of transfusion of blood products was 30.03%. The rate of MOH declined significantly when gestational age increased (OR = 0.940, 95% CI = 0.908 0.974), and was higher in parous women than in nulliparous women (OR = 3.789, 95% CI = 1.769 8.116). Our results showed there was a remarkable connection between gestational age and childbearing history (P = 0.001). The rate of adverse maternal outcomes was higher in women with MOH than those without this history (P < .05). DIC was significantly correlated with MOH (P < .0001) and abdominal hematoma (P < .0001). AUROC of SOFA was significantly higher than that of other scores (P = 0.002) on reoperation. The AUROC of DIC was significantly higher than that of other scores (P = 0.001) on abdominal hematoma. Conclusion: Among women with MNM, gestational age and childbearing history were risk factors for MOH. SOFA scores had relatively good predictive abilities on reoperation, and the DIC scores had relatively good predictive abilities on abdominal hematoma.

The rise in the number of fungal infections is requiring the rapid development of novel antifungal agents. A new polyoxovanadate functionalized by Zn-fluconazole coordination complexes, Zn3(FLC)6V10O28·10H2O (ZnFLC) (FLC = fluconazole) has been synthesized and evaluated for in vitro antifungal against Candida species. The identity of ZnFLC were confirmed by elemental analysis, IR spectrum, and single-crystal X-ray diffraction. The antifungal activities of ZnFLC was screened in 19 Candida species strains using the microdilution checkerboard technique. The minimum inhibitory concentration (MIC80) value of ZnFLC is 4 μg/mL on the azole-resistant clinical isolates of C. albicans HL973, which is lower than the positive control, FLC. The mechanism of ZnFLC against C. albicans HL973 showed that ZnFLC damaged the fungal cell membrane and reduced the ergosterol content. The expression of ERG1, ERG7, ERG11 ERG27, and ERG28, which have effects on the synthesis of ergosterol, were all significantly upregulated by ZnFLC.

BACKGROUND:Because of the deficiency of blood circulation and soft tissue, reconstruction of the eye socket for severe conjunctival sac stenosis in anophthalmic patients is very difficult. In this article, the authors report an innovative technique for conducting a 1-stage operation to reconstruct the contracted eye socket with an autogenic dermal sphere connected to the epidermis (ADSE).METHODS:Five patients, each having a single severely contracted eye socket and conjunctival sac, were included in this study. An ADSE was transplanted into the contracted eye socket and conjunctival sac. After the operation, several observation indexes were evaluated, such as the survival and stability of implanted autologous tissue, the improvement of the contracted eye socket, and the degree of patients' satisfaction with their appearance.RESULTS:All of the implanted epidermal and dermal tissues survived well after 6 months' observation. Although somewhat absorbed, the transplanted dermal ball was plump in the eye socket, and the epithelial tissues merged well with the residual conjunctival epithelium. Moreover, the transplanted epidermis assumed a mucosal appearance in 4 of 5 patients. After reconstruction of the eye socket, the size of the conjunctival sac and depth of the eye socket were corrected sufficiently for patients to wear an ocular prosthesis.CONCLUSIONS:The implantation of an ADSE can reconstruct a severely contracted eye socket in anophthalmic patients. Because of the high survival rate and limited absorption ratio, this 1-stage operation satisfied both patients and their ophthalmologists.

Photodynamic therapy (PDT) is a novel treatment modality that is under intensive preclinical investigations for a variety of diseases, including cancer. Despite extensive studies in this area, selective and effective photodynamic agents that can specifically accumulate in tumors to reach a therapeutic concentration are limited. Although recent attempts have produced photosensitizers (PSs) complexed with various nanomaterials, the tedious preparation steps and poor tumor efficiency of therapy hamper their utilization. Here, we developed a CD44-targeted nanophotodynamic agent by physically encapsulating a photosensitizer, Ce6, into a hyaluronic acid nanoparticle (HANP), which was hereby denoted HANP/Ce6. Its physical features and capability for photodynamic therapy were characterized in vitro and in vivo. Systemic delivery of HANP/Ce6 resulted in its accumulation in a human colon cancer xenograft model. The tumor/muscle ratio reached 3.47 ± 0.46 at 4 h post injection, as confirmed by fluorescence imaging. Tumor growth after HANP/Ce6 treatment with laser irradiation (0.15 W/cm2, 630 nm) was significantly inhibited by 9.61 ± 1.09-fold compared to that in tumor control groups, which showed no change in tumor growth. No apparent systemic and local toxic effects on the mice were observed. HANP/Ce6-mediated tumor growth inhibition was accessed and observed for the first time by 18F-fluoro-2-deoxy-d-glucose positron emission tomography as early as 1 day after treatment and persisted for 14 days within our treatment time window. In sum, our results highlight the imaging properties and therapeutic effects of the novel HANP/Ce6 theranostic nanoparticle for CD44-targeted PDT cancer therapy that may be potentially utilized in the clinic. This HANP system may also be applied for the delivery of other hydrophobic PSs, particularly those that could not be chemically modified.

OBJECTIVE:To analysis the immunological characteristics of patients with severe H1N1 influenza A, and to provide theoretical basis for predicting the prognosis of the disease. METHODS:A retrospective analysis was conducted. The clinical data of 15 patients diagnosed with severe H1N1 influenza A and admitted to Shanghai General Hospital of Nanjing Medical University from October 2015 to December 2016 were collected. All the patients were divided into survival and death groups according to 28-day survival. Clinical characteristics, treatment algorithm, organ function, inflammatory reaction and immune cell status were compared, and Cox regression was used to decide the risk factors of 28-day death in patients with severe H1N1 infection A. RESULTS:All 15 patients with severe H1N1 infection A were enrolled, most of who presented with cough (93.3%), fever (86.7%), sputum production (80.0%), shortness of breath (73.3%), myalgia (40.0%) and fatigue (40.0%). All had been received anti-virus, antibiotics, mechanical ventilation and anti-coagulation therapy; some were treated with prone position, neuromuscular blocker and extracorporeal membrane oxygenation (ECMO). The incidences of acute myocardial and kidney injury were high, and the acute physiology and chronic health evaluation II (APACHE II) score (14.1±6.1) and sequential organ failure assessment (SOFA) score (9.6±4.1) implicated the critical condition. Of 15 patients, 4 patients died in 28 days, while 11 were cured and discharged. Compared with survival group, the patients in death group had higher levels of APACHE II score (22.7±3.8 vs. 11.8±3.8), troponin [cTn (μg/L): 0.52 (0.07, 2.02) vs. 0.15 (0.10, 0.45)] and blood urea nitrogen [BUN (mmol/L): 11.9 (6.7, 29.1) vs. 3.9 (2.7, 6.8)] and a lower level of blood platelets count [PLT (×109/L): 76±33 vs. 146±49, all P < 0.05]. The levels of C-reactive protein (CRP) and interleukin-6 (IL-6) within 24 hours of admission in death group were significantly higher than those of survival group [CRP (mg/L): 172.2±88.5 vs. 74.8±33.1, IL-6 (ng/L): 283.3 (140.1, 711.0) vs. 18.5 (12.7, 71.4), both P < 0.01]. Compared with survival group, the expressions of CD3+, CD4+, CD8+ T cells and natural killer cell (NK cell) in death group were significantly decreased (CD3+ T cell: 0.348±0.119 vs. 0.573±0.106, CD4+ T cell: 0.135±0.046 vs. 0.344±0.098, CD8+ T cell: 0.089±0.057 vs. 0.208±0.054, NK cell: 0.124±0.057 vs. 0.252±0.182, all P < 0.05), but there were no significant differences in CD4+/CD8+ ratio and human leucocyte antigen-DR positive (HLA-DR+) T cell between death group and survival group (CD4+/CD8+ ratio: 1.57±0.26 vs. 1.83±0.54, HLA-DR+ T cell: 0.035±0.022 vs. 0.062±0.036, both P > 0.05). B lymphocyte in death group was significantly higher than that of survival group (0.477±0.136 vs. 0.229±0.121, P < 0.01). Cox regression analysis revealed that APACHE II score [risk ratio (RR) = 20.4, 95% confidence interval (95%CI) = 5.3-31.2, P = 0.017], CD4+ T cell (RR = 11.1, 95%CI = 5.1-20.0, P = 0.048) and CD8+ T cell (RR = 9.1, 95%CI = 4.3-16.7, P = 0.049) were independently risk factors of 28-day survival of patients with severe H1N1 influenza A. CONCLUSIONS:Immunological paralysis and severe inflammatory response were early complicated with severe H1N1 influenza A, and these were significantly associated with prognosis.

Roundabout4 (Robo4) is a gene that is expressed specifically in vasculature and is involved in the angiogenesis and integrity of blood vessels. The expression level of Robo4 increases gradually along with the development of diabetic retinopathy (DR). In this study, we explored the mechanism of transcriptional regulation of Robo4 in retinal endothelial cells, and investigated the effects of this regulation on cellular functions under hyperglycemic conditions. Human retinal endothelial cells (HREC) exposed to hyperglycemia were used to detect the expression levels of specificity protein 1 (SP1) and Robo4 by RT-qPCR and western blotting. Small interfering RNA (SiRNA) transfection technology was used to analyze the regulatory relationship between SP1 and Robo4. The effect of transcription factor SP1 on Robo4 promoter activity and the location of SP1 binding sites were investigated using chromatin immunoprecipitation (ChIP) and luciferase assay. Cell migration, monolayer permeability and tube formation assays were performed to demonstrate the role of SP1/Robo4 in regulating HREC functions in hyperglycemic conditions. The results showed that hyperglycemia upregulated the mRNA and protein levels of SP1 and Robo4 in HREC. Depletion of SP1 by siRNA transfection inhibited the hyperglycemia induced overexpression of Robo4. ChIP combined with luciferase assay showed that under hyperglycemic conditions, SP1 significantly increased the transcriptional level of Robo4 via an additional SP1 binding site at -1912/-1908 in the Robo4 promoter. Repressing the SP1/Robo4 pathway effectively mitigated the abnormity in HREC migration, permeability and angiogenesis induced by hyperglycemia. All these findings indicate that hyperglycemia-induced upregulation of Robo4 is mediated by enhanced transcription of SP1. The SP1/Robo4 signaling pathway can regulate the migratory ability, monolayer permeability and angiogenesis of HREC under hyperglycemic conditions, suggesting that it may play an important role in microvascular dysfunction during DR.

Polyoxometalates (POMs) are inorganic clusters that possess potential anti-bacterial, anti-viral, and anti-tumor activities. Herein, the in vitro anti-proliferation activities of K12[V18O42(H2O)]∙6H2O (V18) have been investigated on the MCF-7 and MDA-MB-231 cell lines. The results indicated that V18 could inhibit the proliferation of MCF-7 (IC50, 11.95 μM at 48 h) in a dose-dependent manner compared to the positive control, 5-fluorouracil (5-Fu, p < 0.05). The anti-proliferation activity of V18 might be mediated by arrest of the MCF-7 cells in the G2/M phase and induction of apoptosis and necrosis. Moreover, V18 can effectively quench the fluorescence of ctDNA. The binding mode between them may be groove or outside stacking binding. V18 can also effectively quench the intrinsic fluorescence of bovine serum albumin (BSA) and human serum albumin (HSA) via static quenching, and changed the conformation of BSA and HSA.