SERPINE2 promotes liver cancer metastasis by inhibiting c-Cbl-mediated EGFR ubiquitination and degradation.

BACKGROUND: Liver cancer is a malignancy with high morbidity and mortality rates. Serpin family E member 2 (SERPINE2) has been reported to play a key role in the metastasis of many tumors. In this study, we aimed to investigate the potential mechanism of SERPINE2 in liver cancer metastasis. METHODS: The Cancer Genome Atlas database (TCGA), including DNA methylation and transcriptome sequencing data, was utilized to identify the crucial oncogene associated with DNA methylation and cancer progression in liver cancer. Data from the TCGA and RNA sequencing for 94 pairs of liver cancer tissues were used to explore the correlation between SERPINE2 expression and clinical parameters of patients. DNA methylation sequencing was used to detect the DNA methylation levels in liver cancer tissues and cells. RNA sequencing, cytokine assays, immunoprecipitation (IP) and mass spectrometry (MS) assays, protein stability assays, and ubiquitination assays were performed to explore the regulatory mechanism of SERPINE2 in liver cancer metastasis. Patient-derived xenografts and tumor organoid models were established to determine the role of SERPINE2 in the treatment of liver cancer using sorafenib. RESULTS: Based on the public database screening, SERPINE2 was identified as a tumor promoter regulated by DNA methylation. SERPINE2 expression was significantly higher in liver cancer tissues and was associated with the dismal prognosis in patients with liver cancer. SERPINE2 promoted liver cancer metastasis by enhancing cell pseudopodia formation, cell adhesion, cancer-associated fibroblast activation, extracellular matrix remodeling, and angiogenesis. IP/MS assays confirmed that SERPINE2 activated epidermal growth factor receptor (EGFR) and its downstream signaling pathways by interacting with EGFR. Mechanistically, SERPINE2 inhibited EGFR ubiquitination and maintained its protein stability by competing with the E3 ubiquitin ligase, c-Cbl. Additionally, EGFR was activated in liver cancer cells after sorafenib treatment, and SERPINE2 knockdown-induced EGFR downregulation significantly enhanced the therapeutic efficacy of sorafenib against liver cancer. Furthermore, we found that SERPINE2 knockdown also had a sensitizing effect on lenvatinib treatment. CONCLUSIONS: SERPINE2 promoted liver cancer metastasis by preventing EGFR degradation via c-Cbl-mediated ubiquitination, suggesting that inhibition of the SERPINE2-EGFR axis may be a potential target for liver cancer treatment.

Cell therapy in end-stage liver disease: replace and remodel.

Liver disease is prevalent worldwide. When it reaches the end stage, mortality rises to 50% or more. Although liver transplantation has emerged as the most efficient treatment for end-stage liver disease, its application has been limited by the scarcity of donor livers. The lack of acceptable donor organs implies that patients are at high risk while waiting for suitable livers. In this scenario, cell therapy has emerged as a promising treatment approach. Most of the time, transplanted cells can replace host hepatocytes and remodel the hepatic microenvironment. For instance, hepatocytes derived from donor livers or stem cells colonize and proliferate in the liver, can replace host hepatocytes, and restore liver function. Other cellular therapy candidates, such as macrophages and mesenchymal stem cells, can remodel the hepatic microenvironment, thereby repairing the damaged liver. In recent years, cell therapy has transitioned from animal research to early human studies. In this review, we will discuss cell therapy in end-stage liver disease treatment, especially focusing on various cell types utilized for cell transplantation, and elucidate the processes involved. Furthermore, we will also summarize the practical obstacles of cell therapy and offer potential solutions.

[Temporal and Spatial Characteristics of Net CO2 Emissions and Decoupling Analysis in Yangtze River Economic Belt].

Calculating the fossil energy consumption, revealing the temporal and spatial evolution characteristics of net CO2 emissions, and analyzing the decoupling effect between social development and net CO2 emissions in different regions of the Yangtze River Economic Belt (YREB) is crucial to support the different regions, allowing them to select their individual industrial development and carbon emission reduction path. The results showed that:① from 1999 to 2012, YREB became greener, the CO2 emission of the YREB increased by 2244.23 million tons, and the carbon sink increased by 148.07 million tons during the research period. ② From 2013 to 2018, the area of medium-high carbon sequestration (NPP>800 g·m-2, count for C) increased by 23.25%, compared with that from 1999-2012. ③ A highly decoupling effect between social development and net CO2 emissions was found in the downstream of the YREB. The highest decoupling cities in the upstream, midstream, and downstream accounted for 12%, 34%, and 54% of the highest decoupling cities in the YREB, respectively.

Detectable 2019-nCoV viral RNA in blood is a strong indicator for the further clinical severity.

The novel coronavirus (2019-nCoV) infection caused pneumonia. we retrospectively analyzed the virus presence in the pharyngeal swab, blood, and the anal swab detected by real-time PCR in the clinical lab. Unexpectedly, the 2109-nCoV RNA was readily detected in the blood (6 of 57 patients) and the anal swabs (11 of 28 patients). Importantly, all of the 6 patients with detectable viral RNA in the blood cohort progressed to severe symptom stage, indicating a strong correlation of serum viral RNA with the disease severity (p-value = 0.0001). Meanwhile, 8 of the 11 patients with annal swab virus-positive was in severe clinical stage. However, the concentration of viral RNA in the anal swab (Ct value = 24 + 39) was higher than in the blood (Ct value = 34 + 39) from patient 2, suggesting that the virus might replicate in the digestive tract. Altogether, our results confirmed the presence of virus RNA in extra-pulmonary sites.

Note on sediment removal efficiency in oil-grit separators.

Oil-grit separators (OGSs) are one type of best management practice, designed to remove oil and grit from stormwater runoff (e.g., from parking lots and paved roads). This note examines scaling parameters for OGS removal efficiency. Three dimensionless parameters are chosen as scaling parameters: Hazen number (Ha), Reynolds number (Re) and Froude number (Fr). The Hazen number is a ratio of hydraulic residence time to particle settling time. The Reynolds number measures the surrounding turbulence effects on sediment removal efficiency. The Froude number represents the ratio of inertial and gravitational forces, which indicates the influence of gravity on fluid motion. The collected data from the literature on sediment removal in OGSs can be represented by a single curve when the Hazen, Reynolds, and Froude numbers are combined into a new scaling parameter (HRF = Ha(Re/Fr)). A general form is proposed to correlate the sediment removal efficiency with this new parameter. This generalized prediction method can be used as a preliminary performance indicator for OGS units. The obtained curve can also be used to adjust raw laboratory and field measurement data to improve the evaluation of the performance of various OGSs.

Experimental study of hydraulics and sediment capture efficiency in catchbasins.

Catchbasins (also known as gully pot in the UK and Australia) are used to receive surface runoff and drain the stormwater into storm sewers. The recent interest in catchbasins is to improve their effectiveness in removing sediments in stormwater. An experimental study was conducted to examine the hydraulic features and sediment capture efficiency in catchbasins, with and without a bottom sump. A sump basin is found to increase the sediment capture efficiency significantly. The effect of inlet control devices, which are commonly used to control the amount of flow into the downstream storm sewer system, is also studied. These devices will increase the water depth in the catchbasin and increase the sediment capture efficiency. Equations are developed for predicting the sediment capture efficiency in catchbasins.

Serum levels of soluble vascular cell adhesion molecules may correlate with the severity of dengue virus-1 infection in adults.

The damage of vascular endothelial cells has been speculated to be involved in the pathogenesis of dengue virus (DENV) infection. However, little is known about the role of soluble vascular cell adhesion molecule-1 (sVCAM-1) in predicting the severity of dengue infection in adults. In this study, 51 adults with DENV-1 infection (21 with severe dengue and 30 with dengue fever (DF) were included, and their serum levels of sVCAM-1 and other parameters were determined. The results indicated that the levels of sVCAM-1 were elevated on days 1-3 to 16.75 (11.55-34.74) ng/mL in the severe dengue patients. These levels increased rapidly to peak values of 43.53 (37.15-47.02) ng/mL on days 10-12 and then declined; however, the values were maintained at a high level (38.07 (26.06-39.63) ng/mL). Other parameters, including reduced platelet (PLT) counts, neutrophil (NEU) counts and increased levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK), were also observed in the severe dengue group but not in the DF group. The levels of cytokines, such as interleukin (IL)-6, IL-10, interferon γ (IFNγ), and tumor necrosis factor α (TNF-α), were transiently increased in the severe dengue patients. Among the aforementioned parameters, only sVCAM-1 levels were significantly elevated earlier and more persistently in the severe dengue patients than in the DF patients. sVCAM-1 positively correlated with the levels of ALT, AST, LDH, TNF-α, and IL-6 and negatively correlated with the levels of PLT, NEU, and viremia. Notably, the high levels of sVCAM-1 were closely associated with the severe dengue patients. In conclusion, sVCAM-1 may be a superior indicator for monitoring the severity of dengue.

[Analysis of hepatitis C virus (HCV) subtypes in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong province].

OBJECTIVE: To explore the transmission routes, genotypes/subtypes distribution and genetic character of HCV in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province. METHODS: Reverse transcription (RT) nested PCR was performed to amplify the HCV NS5B gene region from 95 HIV/HCV co-infected and 99 HCV mono-infected individuals lived in Guangdong province. The PCR products were then sequenced for HCV subtyping. Genetic analysis was done by MEGA4 software. RESULTS: (1) HIV/HCV co-infected individuals infected HCV mostly through injection drug use (IDU, 78.9%), the HCV subtypes were identified as 6a (53.7%), 3a (17.9%), 1b (15.8%), 3b (11.6%) and 1a (1.0%) respectively, the genetic distance within subtype 1b was longer than those within other subtypes, the predominant HCV subtype in HIV/HCV co-infected individuals infected through IDU was 6a (60.0%). (2) HCV mono-infected individuals infected HCV mostly through blood or blood products transfusions (80.8%), the HCV subtypes were identified as 1b (67.7%), 6a (17.2%), 3a (6.1%), 2a (5.0%), 3b (2.0%), 4a (1.0%) and 5a (1.0%) respectively, the genetic distance within subtype 1b was also longer than those within other subtypes, the predominant HCV subtype in HCV mono-infected individuals infected through blood or blood products transfusions was 1b (76.2%). CONCLUSION: The diversity of HCV subtypes in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province was high, both the major transmission route and HCV subtype between HIV/HCV co-infected individuals and HCV mono-infected individuals were different.

Both viremia and cytokine levels associate with the lack of severe disease in secondary dengue 1 infection among adult Chinese patients.

Secondary dengue infections are frequently associated with increased risk for dengue hemorrhagic fever and dengue shock syndrome. Surprisingly, we observed no dengue hemorrhagic fever cases among 353 hospitalized dengue-infected patients including 212 with primary, and 141 with secondary dengue 1 infection in China. To explore virological and immunological mechanisms which may account for this unexpected clinical observation, we assessed dengue viremia, type I interferon and inflammatory cytokine levels in these patients. While the levels of viremia and inflammatory cytokines are indistinguishable between primary and secondary infections, IFNα levels are significantly higher in primary than that in secondary infection. However, IFNα levels are positively correlated with dengue viremia levels (p<0.0001), but negatively correlated with the platelet counts (p<0.0001) and serum ALT levels (p = 0.0003). These results provide direct in vivo evidence that clinical dengue disease severity is affected by both viral and human immune factors.

Unique impacts of HBV co-infection on clinical and laboratory findings in a recent dengue outbreak in China.

High prevalence of hepatitis B virus (HBV) infection in China offers a unique setting to examine HBV's influence on the presentation of dengue fever. In 398 patients admitted for suspected dengue fever, 89% (353/398) were positive for dengue IgM antibodies. Among dengue-infected patients, 8% (29/353) had chronic HBV co-infection. Only dengue virus serotype 1 was identified by virus isolation and reverse transcriptase-polymerase chain reaction assays. No case of dengue hemorrhagic fever/dengue shock syndrome was diagnosed. In addition to routine clinical tests, interleukin 2 (IL-2), IL-4, IL-6, IL-10, interferon gamma (IFNgamma), and tumor necrosis factor alpha (TNFalpha) levels were measured in the sera of 95% (334/353) of dengue-infected subjects as well as controls. Surprisingly, HBV/dengue co-infected patients made less IL-6 (P < 0.05) and TNFalpha (P < 0.05) than patients with only dengue infection. Similar levels of IL-4, IL-10, and IFNgamma were found in both groups. Thus, HBV co-infection seems to alter the cytokine production pattern when patients contract dengue infection.

[Immunodominance in CD8+ T cell responses to HIV-1 synthesized epitopes].

OBJECTIVE: To investigate immunodominance in CD8+ T cell responses to human immunodeficiency virus type 1 (HIV-1) epitopes. METHODS: Frequency of Interferon-gamma (IFN-gamma) secreting cells and the proliferation percentage of CD8+ T cells in PBMC from an HIV-1-infected long term nonprogressor (LTNP) were assessed after stimulation with either the 34 pools of 701 overlapping peptides covering the regions of HIV-1 Env, Pol, Gag, Vif, Nef, Tat or some single peptides, by using various assays including enzyme-linked immunospot (ELISPOT) and CFSE Carboxy-fluorescein diacetate, succinimidyl ester (CFSE) labeling and flow cytometry. RESULTS: HIV-1 Gag peptides induced the highest frequency of IFN-gamma secreting cells, followed by Nef, Tat, and Vif. Meanwhile, Env and Pol failed to induce significant responses. In the IFN-gamma ELISPOT assay, stimulation with single peptide and the corresponsive peptide pool generated analogous results. In addition, the frequencies of IFN-gamma secreting cells and the proliferation percentage of CD8+ T cells detected-ELISPOT and CFSE labeling and flow cytometry were proportional, when single peptides were used for stimulation. CONCLUSION: CD8+ T cells can respond to some specific HIV-1 epitopes and induce immunodominant responses. As a complimentary approach to the standard of ELISPOT assay, We recommend a novel CFSE labeling and flow cytometry assay for the examination of immunodominance in studies of HIV-1 specific proliferation percentage of CD8+ T cell responses.