LncRNA PWRN1 inhibits the progression of hepatocellular carcinoma by activating PKM2 activity.

Hepatocellular carcinoma (HCC) is one of the most prevalent and leading causes of cancer-related mortality worldwide. Long non-coding RNAs (lncRNAs) have been demonstrated to play vital roles in cancer development and progression. The lncRNA PWRN1 (PWRN1), acts as a tumor suppressor factor, which is low expressed in some cancers. However, the molecular mechanisms underlying the effects of PWRN1, especially the regulatory relationship with RNA binding protein in HCC remain largely unknown. In the present study, we demonstrated that PWRN1 was significantly down-regulated in HCC and correlated with better prognosis; furthermore, gain-of-function experiments showed that PWRN1 inhibited the proliferation of HCC cells. We further found that PWRN1 up-regulated pyruvate kinase activity and thus hinders the proliferation of HCC in vitro and in vivo. Mechanistically, pyruvate kinase M2 (PKM2) was bound to it and maintained the high activity state of PKM2, thereby hindering PKM2 from entering the nucleus in the form of low-activity dimers, reducing the expression of c-Myc downstream gene LDHA, leading to a decrease in lactate levels, and inhibiting the growth of tumor cells. In addition, PWRN1 was found to inhibit aerobic glycolysis. Finally, TEPP-46, a pyruvate kinase activator, appeared to inhibit HCC proliferation by maintaining tetramer stability and increasing pyruvate kinase activity. Taken together, our results provide new insights into the biology hindering HCC proliferation and indicate that PWRN1 in combination with PKM2 activators might represent a novel therapeutic target for HCC.

LncRNA CRNDE Drives the Progression of Hepatocellular Carcinoma by inducing the Immunosuppressive Niche.

As a crucial protumorigenic long noncoding RNA, colorectal tumor differential expression (CRNDE) has been confirmed to facilitate the progression of various cancers. However, its role in the tumor microenvironment (TME) of hepatocellular carcinoma (HCC) is still unclear. Here we determined that CRNDE was upregulated in HCC samples and that CRNDE-positive cells were predominantly enriched in malignant tumor cells. In vivo functional assays revealed that CRNDE-induced tumor cells supported HCC progression, recruited abundant granulocyte myeloid-derived suppressor cells (G-MDSCs) and restricted the infiltration of T cells. In terms of mechanisms, CRNDE bound with Toll-like receptor 3 (TLR3) and activated NF-κB signaling to increase the secretion of c-x-c motif chemokine ligand 3 (CXCL3). CRNDE knockdown could significantly suppress the accumulation of G-MDSCs and enhance the infiltration of T cells in the TME of HCC in vivo. Taken together, our study reveals the CRNDE-NF-κB-CXCL3 axis plays a crucial role in driving the immunosuppressive niche to facilitate HCC progression by recruiting G-MDSCs.

N-glycosylation stabilizes MerTK and promotes hepatocellular carcinoma tumor growth.

Despite the evidences of elevated expression of Mer tyrosine kinase (MerTK) in multiple human cancers, mechanisms underlying the oncogenic roles of MerTK in hepatocellular carcinoma (HCC) remains undefined. We explored the functional effects of MerTK and N-Glycosylated MerTK on HCC cell survival and tumor growth. Here, we show that MerTK ablation increases reactive oxygen species (ROS) production and promotes the switching from glycolytic metabolism to oxidative phosphorylation in HCC cells, thus suppressing HCC cell proliferation and tumor growth. MerTK is N-glycosylated in HCC cells at asparagine 294 and 454 that stabilizes MerTK to promote oncogenic transformation. Moreover, we observed that nuclear located non-glycosylated MerTK is indispensable for survival of HCC cells under stress. Pathologically, tissue microarray (TMA) data indicate that MerTK is a pivotal prognostic factor for HCC. Our data strongly support the roles of MerTK N-glycosylation in HCC tumorigenesis and suggesting N-glycosylation inhibition as a potential HCC therapeutic strategy.

Neutrophil-to-Lymphocyte Ratios Are Closely Associated With the Severity and Course of Non-mild COVID-19.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is spreading worldwide. Measuring the prevention and control of the disease has become a matter requiring urgent focus. Objective: Based on coronavirus disease 2019 (COVID-19) clinical data from Wuhan, we conducted an in-depth analysis to clarify some of the pathological mechanisms of the disease and identify simple measures to predict its severity early on. Methods: A total of 230 patients with non-mild COVID-19 were recruited, and information on their clinical characteristics, inflammatory cytokines, and T lymphocyte subsets was collected. Risk factors for severity were analyzed by binary logistic regression, and the associations of neutrophil-to-lymphocyte ratios (N/LRs) with illness severity, disease course, CT grading, inflammatory cytokines, and T lymphocyte subsets were evaluated. Results: Our results showed that the N/LRs were closely related to interleukin (IL)-6 and IL-10 (P < 0.001, P = 0.024) and to CD3+ and CD8+ T lymphocytes (P < 0.001, P = 0.046). In particular, the N/LRs were positively correlated with the severity and course of the disease (P = 0.021, P < 0.001). Compared to the values at the first test after admission, IL-6 and IL-10 were significantly decreased and increased, respectively, as of the last test before discharge (P = 0.006, P < 0.001). More importantly, through binary logistic regression, we found that male sex, underlying diseases (such as cardiovascular disease), pulse, and N/LRs were all closely related to the severity of the disease (P = 0.004, P = 0.012, P = 0.013, P = 0.028). Conclusions: As a quick and convenient marker of inflammation, N/LRs may predict the disease course and severity level of non-mild COVID-19; male sex, cardiovascular disease, and pulse are also risk factors for the severity of non-mild COVID-19.

[Value of neutrophil-to-lymphocyte ratio in the classification diagnosis of coronavirus disease 2019].

OBJECTIVE: To investigate the clinical significance of neutrophil-to-lymphocyte ratio (NLR) in classification of patients with coronavirus disease 2019 (COVID-19). METHODS: A retrospective analysis was performed on 72 patients with COVID-19 admitted to the critical ward of Cancer Center of Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology in Wuhan from February to March in 2020. The patients were divided into two groups: moderate type (non-severe group) and severe/critical type (severe group). The results of white blood cell count (WBC), neutrophil count (NEU), lymphocyte count (LYM), interleukin-6 (IL-6) and D-dimer were collected at the 2nd day after admission from the two groups, and the NLR was calculated. The diagnostic value of WBC, NEU, LYM, IL-6, D-dimer and NLR on COVID-19 classification was evaluated by the receiver operating characteristic (ROC) curve. RESULTS: A total of 72 COVID-19 patients were enrolled, among whom 52 were moderate, 17 were severe, and 3 were critical. The most common clinical manifestations of patients were fever (70.8%), cough (36.1%), chest tightness and breathlessness (37.5%), diarrhea (15.3%), fatigue (15.3%), vomiting and nausea (11.1%), occasionally accompanied by acute dyspnea (2.8%), and only one patient had no clinical symptom (1.4%). The levels of WBC, NEU, IL-6, D-dimer and NLR in the severe group were significantly higher than those in the non-severe group [WBC (×109/L): 7.81±3.65 vs. 5.34±1.69, NEU (×109/L): 5.83±3.13 vs. 3.24±1.53, IL-6 (ng/L): 133.63 (71.09, 249.61) vs. 28.05 (6.41, 101.24), D-dimer (mg/L): 0.86 (0.31, 2.56) vs. 0.33 (0.20, 0.71), NLR: 6.14±4.75 vs. 2.66±1.93, all P < 0.05], and the level of LYM was significantly lower than that in the non-severe group (×109/L: 1.09±0.56 vs. 1.49±0.74, P < 0.05). The results of ROC curve analysis showed that the areas under ROC curve (AUC) of WBC, NEU, LYM, IL-6, D-dimer and NLR for COVID-19 classification were 0.790 [95% confidence interval (95%CI) was 0.684-0.897), 0.869 (95%CI was 0.789-0.949), 0.719 (95%CI was 0.592-0.847), 0.790 (95%CI was 0.682-0.898), 0.676 (95%CI was 0.526-0.827), and 0.888 (95%CI was 0.814-0.963) respectively. The AUC of NLR was the highest, which was of high diagnostic value; when the optimum cut-off value of NLR was 3.00, the sensitivity was 100%, and the specificity was 73.1%. CONCLUSIONS: NLR can be used as a biomarker to predict classification of COVID-19 patients independently, which can provide a theoretical basis for the classification management of COVID-19 patients.

Risk-adapted Treatment Strategy For COVID-19 Patients.

BACKGROUND: There are no clear expert consensus or guidelines on how to treat 2019 coronavirus disease (COVID-19). The objective of this study is to investigate the short-term effect of risk-adapted treatment strategy on patients with COVID-19. METHODS: We collected the medical records of 55 COVID-19 patients for analysis. We divided these patients into mild, moderate and severe groups, and risk-adapted treatment approaches were given according to the illness severity. RESULTS: Twelve patients were in mild group and 22 were in moderate group (non-severe group, n=34), and 21 patients were in severe group. At the end of the first two weeks after admission, clinical manifestations had completely despeared in 31(91.2%)patients in non-severe group, and 18(85.7%) patients in severe group (p=0.85). Both groups had a satisfied chest CT imaging recovery, which includes 22(64.7%) patients in non-severe group and 12(57.1%) patients in severe group recovered at least 50% of the whole leisions in the first week, and 28(82.4%) and 16(76.2%) recovered at least 75% in the second week, respectively. There were no significant differences in SARS-CoV-2 nucleic acid negativity (p=0.92). There were also no significant differences in the levels of SARS-CoV-2-IgM and IgG antibody production between the two groups (p=0.13, 0.62). There were 45 cases were discharged from the hospital, and no patients died at the time of this clinical analysis. CONCLUSIONS: Risk-adapted treatment strategy was associated with significant clinical manifestations alleviation and clinical imaging recovery. In severe COVID-19 patients, early and short-term use of low-dose methylprednisolone was beneficial and did not delay SARS-CoV-2 nucleic acid clearance and influence IgG antibody production.

Human encephalitis caused by pseudorabies virus infection: a case report.

Pseudorabies virus (PRV) primarily infects swine but can infect cattle, dogs, and cats. Several studies have reported that PRV can cross the specie barrier and induce human encephalitis, but a definitive diagnosis of human PRV encephalitis is debatable due to the lack of PRV DNA detection. Here, we report a case of human PRV encephalitis diagnosed by the next-generation sequencing (NGS) of PRV sequences in the cerebrospinal fluid (CSF) of a patient. A male pork vendor developed fever and seizures for 6 days. NGS results showed PRV sequences in his CSF and blood. Sanger sequencing showed that PRV DNA in the CSF and PRV antibodies in both the CSF and blood were positive. MRI results revealed multiple inflammatory lesions in the bilateral hemisphere. Based on the clinical and laboratory data, we diagnosed the patient with PRV encephalitis. This case suggests that PRV can infect humans, causing severe viral encephalitis. People at risk of PRV infection should improve their self-protection awareness.

Artemisinin suppresses hepatocellular carcinoma cell growth, migration and invasion by targeting cellular bioenergetics and Hippo-YAP signaling.

The primary liver cancer (PLC) is one of the leading causes of cancer-related death worldwide. The predominant form of PLC is hepatocellular carcinoma (HCC), which accounts for about 85% of all PLC. Artemisinin (ART) was clinically used as anti-malarial agents. Recently, it was demonstrated to inhibit cell growth and migration in multiple cancer types. However, the molecular mechanism underlying these anti-cancer activity remains largely unknown. Herein, it is discovered that ART dramatically suppresses HCC cell growth in vitro through arresting cell cycle progression, and represses cell migration and invasion via regulating N-cadherin-Snail-E-cadherin axis. In addition, the disruption of cellular bioenergetics contributed to ART-caused cell growth, migration and invasion inhibition. Moreover, ART (100 mg/kg, intraperitoneally) substantially inhibits HCC xenograft growth in vivo. Importantly, Hippo-YAP signal transduction is remarkably inactivated in HCC cells upon ART administration. Collectively, these data reveal a novel mechanism of ART in regulating HCC cell growth, migration, and invasion, which indicates that ART could be considered as a potential drug for the treatment of HCC.

[Influence of hypocalcemia on the prognosis of septic patients].

OBJECTIVE: To investigate the effect of serum calcium level on the prognosis of patients with sepsis. METHODS: Clinical data of 119 patients with sepsis admitted to intensive care medicine (ICU) of the First Affiliated Hospital of the University of Science and Technology of China from January 2017 to October 2018 were retrospectively analyzed. Gender, age, and C-reactive protein (CRP), procalcitonin (PCT), serum calcium levels, acute physiology and chronic health evaluation II (APACHE II), sequential organ failure score (SOFA) within 24 hours of diagnosis, and 28-day mortality were collected. The patients were divided into the normal serum calcium group (serum calcium 2.00-2.67 mmol/L) and the hypocalcemia group (serum calcium < 2.00 mmol/L) according to their serum calcium level. The patients were divided into survival group and death group according to 28-day prognosis. Pearson correlation test was used to analyze the correlation between serum calcium level and clinical indicators. Receiver operator characteristic (ROC) curve was used to analyze the predictive value of serum calcium level on prognosis. RESULTS: A total of 119 patients with sepsis were included, including 50 patients with normal serum calcium, with serum calcium level of (2.14±0.10) mmol/L; and 69 patients of hypocalcemia, and the incidence of hypocalcemia was 57.98%, with serum calcium level of (1.81±0.14) mmol/L. In the hypocalcemia group, except that the APACHE II score was significantly higher than that of the normal serum calcium group (25.59±5.52 vs. 22.28±4.89, P < 0.01), there was no significant difference in gender, age, CRP, PCT and SOFA score between the two groups. The 28-day mortality rate of the hypocalcemia group was significantly higher than that of the normal serum calcium group [78.26% (54/69) vs. 48.00% (24/50), χ2 = 10.45, P < 0.01]. The level of serum calcium in the death group was significantly lower than that in the survival group (mmol/L: 1.90±0.20 vs. 2.04±0.19), while the APACHE II score was significantly higher than the survival group (25.78±5.25 vs. 21.20±4.68), with statistically significant differences (both P < 0.01). There was a negative correlation between serum calcium level and PCT, APACHE II scores in patients with sepsis (r1 = -2.10, P1 = 0.04; r2 = -3.91, P2 < 0.01), but no correlation with CRP and SOFA score (r1 = 0.75, P1 = 0.46; r2 = -1.21, P2 = 0.23). The ROC curve analysis showed that the area under the ROC curve (AUC) for predicting the prognosis of sepsis patients with serum calcium level was 0.70 [95% confidence interval (95%CI) = 0.602-0.798], and the best cut-off value was 1.92 mmol/L, with the sensitivity was 52.56%, and the specificity was 82.93%. CONCLUSIONS: The prognosis of sepsis patients with hypocalcemia is poor. Serum calcium level can be used as a predictor of prognosis in patients with sepsis.

Polybenzimidazole/Mxene composite membranes for intermediate temperature polymer electrolyte membrane fuel cells.

This report demonstrated the first study on the use of a new 2D nanomaterial (Mxene) for enhancing membrane performance of intermediate temperature (>100 °C) polymer electrolyte membrane fuel cells (ITPEMFCs). In this study, a typical Ti3C2T x -MXene was synthesized and incorporated into polybenzimidazole (PBI)-based membranes by using a solution blending method. The composite membrane with 3 wt% Ti3C2T x -MXene showed the proton conductivity more than 2 times higher than that of pristine PBI membrane at the temperature range of 100 °C-170 °C, and led to substantial increase in maximum power density of fuel cells by ∼30% tested at 150 °C. The addition of Ti3C2T x -MXene also improved the mechanical properties and thermal stability of PBI membranes. At 3 wt% Ti3C2T x -MXene, the elongation at break of phosphoric acid doped PBI remained unaffected at 150 °C, and the tensile strength and Young's modulus was increased by ∼150% and ∼160%, respectively. This study pointed out promising application of MXene in ITPEMFCs.

[Effect of albumin and hemoglobin level on prognosis of patients with uncomplicated severe traumatic brain injury: a retrospective cohort study].

OBJECTIVE: To evaluate the best level of albumin and hemoglobin for the patients with uncomplicated severe traumatic brain injury (TBI). METHODS: A retrospective cohort study was conducted. One hundred and sixty-eight patients with uncomplicated severe TBI admitted to intensive care unit (ICU) of Anhui Provincial Hospital were enrolled. The relationship between albumin and hemoglobin level within 3 days after admission and prognosis was analyzed. Mean 3-day albumin level was obtained, and then the patients were divided into <25, 25-28, 29-31 and ≥32 g/L groups according to quartiles based on mean albumin, and also were divided into <90, 90-99, 100-109 and≥110 g/L groups according to the mean hemoglobin concentration. Multivariable log-binomial regression was used to model the association between mean albumin and hemoglobin concentration and prognosis. RESULTS: One hundred and nine patients were enrolled based on inclusion/exclusion criteria. Among them, 32 patients (29.4%) received a red blood cell (RBC) transfusion, and 24 patients (22.0%) were given albumin treatment. According to the average level of albumin, there were significant differences in mortality among <25, 25-28, 29-31 and ≥32 g/L groups [85.2%(23/27), 59.3%(16/27), 32.1%(9/28), 44.4%(12/27), respectively, P=0.001]. According to the hemoglobin level, there was no significant difference in mortality rate among <90, 90-99, 100-109 and ≥110 g/L groups[61.8%(34/55), 43.8%(7/16), 53.3%(8/15), 47.8%(11/23), respectively, P>0.05]. When using the albumin level, Glasgow coma score (GCS), age and time of onset for logistic analysis, albumin level and age had influence on the mortality of patients, and mortality rate was generally decreased with an increase in albumin. But when the levels of albumin was ≥32 g/L, the risk of death was higher than in the 29-31 g/L group[relative risk (RR) of albumin 29-31 g/L=0.070, 95% confidence interval (95%CI) 0.015-0.331, P=0.001; RR of albumin≥32 g/L=0.153, 95%CI 0.035-0.663, P=0.012; RR of age=0.691, 95%CI 0.526-0.907, P=0.008]. When the hemoglobin level, GCS, age and time of onset were used for logistic regression analysis, only GCS entered the regression model (RR=0.696,95%CI 0.550-0.880, P=0.002), illustrating that the hemoglobin level had no significant influence on mortality, and mortality rate declined with an increase in GCS. CONCLUSIONS: The most favorable level of albumin for uncomplicated severe traumatic brain injury is 29-31 g/L. There is no difference in mortality rate when hemoglobin >90 g/L.

Analysis of S. Epidermidis icaA and icaD genes by polymerase chain reaction and slime production: a case control study.

BACKGROUND: Staphylococcus epidermidis is a common pathogen in medical device-associated infections and have an ability to form adherent slime. We aimed to study the effects of icaA and icaD genes on the slime formation of Staphylococcus epidermidis associated with catheter-associated infections. METHODS: S. epidermidis isolates from the central venous catheter blood of patients with catheter-associated infections, and from the nasal vestibules of healthy volunteers, intensive care unit hospital staff, and patients, were collected. Slime phenotype was determined by Congo red agar test. The icaA/D was detected by polymerase chain reaction. Slime was examined using scanning electron microscopy. RESULTS: A total of 82 S. epidermidis isolates were collected. We found a statistically significant difference with regards to slime production between the clinical isolates from the catheter blood specimens and those from the nasal vestibules (p<0.05). All S. epidermidis slime positive strains isolated were icaA positive. There was a greater correlation between the presence of both icaA and icaD and the slime production than the single expression of icaA or icaD and the presence of slime in all groups. The co-expression of mecA and icaD was associated with enhanced resistance to antibiotics. CONCLUSION: S. epidermidis bacteria are significant nosocomial pathogens, and icaA/D can clarify the adhesion mechanism in the pathogenesis of infections associated with medical devices. This study result could be useful for the development of rapid diagnosis for slime producing and methicillin resistant S. epidermidis strains.