Kinetic changes in virology, specific antibody response and imaging during the clinical course of COVID-19: a descriptive study.

BACKGROUND: To explore the kinetic changes in virology, specific antibody response and imaging during the clinical course of COVID-19. METHODS: This observational study enrolled 20 patients with COVID-19, who were hospitalized between January 20-April 6, 2020, in the two COVID-19 designated hospitals of Zhoushan, Zhejiang and Rushan, Shandong, China, The laboratory findings, imaging, serum response to viral infection, and viral RNA level in the throat and stool samples were assessed from onset to recovery phase in patients with COVID-19. RESULTS: SARS-COV-2 RNA was positive as early as day four. It remained positive until day 55 post-onset in the sputum-throat swabs and became negative in most cases (55%) within 14 days after onset. Lymphocytopenia occurred in 40% (8/20) of patients during the peak infection period and returned to normal at week five. The most severe inflammation in the lungs appeared in week 2 or 3 after onset, and this was completely absorbed between week 6 and 8 in 85.7% of patients. All patients had detectable antibodies to the receptor binding domain (RBD), and 95% of these patients had IgG to viral N proteins. The antibody titer peaked at week four. Anti-S IgM was positive in 7 of 20 patients after week three. CONCLUSIONS: All COVID-19 patients in this study were self-limiting and recovered well though it may take as long as 6-8 weeks. Our findings on the kinetic changes in imaging, serum response to viral infection and viral RNA level may help understand pathogenesis and define clinical course of COVID-19.

SPINK1 promotes cell growth and metastasis of lung adenocarcinoma and acts as a novel prognostic biomarker.

Serine protease inhibitor Kazal type 1 (SPINK1) plays a role in protecting the pancreas against premature activation of trypsinogen and is involved in cancer progression. SPINK1 promoted LAC cells growth, migration, and invasion. Mechanistically, we found that SPINK1 promoted LAC cells migration and invasion via up-regulating matrix metalloproteinase 12 (MMP12). We observed that SPINK1 expression was only up-regulated in lung adenocarcinoma (LAC) tissues, and was an independent prognostic factor for poor survival. Our results indicate that SPINK1 might be a potential biomarker for LAC that promotes progression by MMP12. [BMB Reports 2018; 51(12): 648-653].

Sulfuration of an Fe-N-C Catalyst Containing Fex C/Fe Species to Enhance the Catalysis of Oxygen Reduction in Acidic Media and for Use in Flexible Zn-Air Batteries.

During the preparation of atomically dispersed Fe-N-C catalysts, it is difficult to avoid the formation of iron-carbide-containing iron clusters ("Fex C/Fe"), along with the desired carbon matrix containing dispersed FeNx sites. As a result, an uncertain amount of the oxygen reduction reaction (ORR) occurs, making it difficult to maximize the catalytic efficiency. Herein, sulfuration is used to boost the activity of Fex C/Fe, forming an improved system, "FeNC-S-Fex C/Fe", for catalysis involving oxygen. Various spectroscopic techniques are used to define the composition of the active sites, which include Fe-S bonds at the interface of the now-S-doped carbon matrix and the Fex C/Fe clusters. In addition to outstanding activity in basic media, FeNC-S-Fex C/Fe exhibits improved ORR activity and durability in acidic media; its half-wave potential of 0.821 V outperforms the commercial Pt/C catalyst (20%), and its activity does not decay even after 10 000 cycles. Interestingly, the catalytic activity for the oxygen evolution reaction (OER) simultaneously improves. Thus, FeNC-S-Fex C/Fe can be used as a high-performance bifunctional catalyst in Zn-air batteries. Theoretical calculations and control experiments show that the original FeNx active centers are enhanced by the Fex C/Fe clusters and the Fe-S and C-S-C bonds.

Increased expression of T cell immunoglobulin and mucin domain 4 is positively associated with the disease severity of patients with ankylosing spondylitis.

Monocytes and associated cytokines have been shown to be involved in the pathogenesis of ankylosing spondylitis (AS). T cell immunoglobulin and mucin domain 4 (Tim-4) was identified on monocytes/macrophages and dentritic cells (DCs) and plays important roles in regulating the activities of macrophages and DCs. The current study investigated the association between Tim-4 expression and AS. Our results showed that Tim-4 expression on monocytes and Tim-4 level in plasma were highly increased in AS patients than in controls. Furthermore, TNF-α production and bath ankylosing spondylitis disease activity index (BASDAI) have positive relationships with Tim-4 expression in AS patients. High expression of Tim-4 was thought to contribute to the pathogenesis and an underlying mechanism of AS.

Anticancer effects of cinnamic acid in lung adenocarcinoma cell line h1299-derived stem-like cells.

Lung cancer is a lethal solid tumor with poor prognosis because of its high metastasis and resistance to current therapies. Recently, cancer stem cells (CSCs) were suggested to be major contributors to tumorigenicity and cancer relapse. However, therapeutic targets for lung cancer-related CSCs remain undetermined. The objective of the current study was to investigate whether cinnamic acid (CINN) exerts an antitumor activity against sphere-derived lung CSCs. In this study, CSCs were isolated from the non-small cell lung cancer cell line H1299 as tumor spheres under CSC-selective conditions, and found to have increased tumorigenicity, chemoresistance, and higher expression of both embryonic stem cell-related and drug resistance-related genes compared with parental cells. These observations are consistent with the notion that CSCs are tumorigenic, display the ability to self-renew, and generate differentiated progeny that constitute the majority of cells in tumors. Treatment of sphere-derived stem cells with CINN could diminish their CSC-like abilities by decreasing their proliferation and invasive abilities and facilitating their differentiation into CD133-negative cells. Furthermore, CINN treatment increased the sensitivity of CSCs to chemotherapeutic drugs through apoptosis. Of note, xenotransplantation experiments revealed that CINN combined with cisplatin had a synergistic effect in inhibiting the tumorigenicity of CSCs. In summary, our study clearly revealed the presence of a population of sphere-forming cells with stem-like properties among H1299 cells and CINN can attenuate CSC properties of this stem-like cell population. The potential of CINN should be verified further in future studies of anti-CSC therapy.

Chan-Yu-Bao-Yuan-Tang, the water extract of a chinese medicine prescription, induces s-phase arrest and mitochondria-mediated apoptosis in human lung adenocarcinoma cells.

Previous clinical studies have shown good efficacy of the traditional Chinese medicinal herbal water extract Chan-Yu-Bao-Yuan-Tang (CYBYT) in lung cancer patients. In this study, CYBYT's effects on proliferation and apoptosis of human lung adenocarcinoma cell line SPC-A-1 cultured in vitro were explored. An XTT assay, cell cycle analysis, Annexin V-FITC staining and Western blot were applied to identify the viability of cells, cell cycle arrest, stages of apoptosis, and signaling proteins, respectively. The results showed that CYBYT inhibited the growth of SPC-A-1 cells by reducing the cells in G0/G1 phase but increasing them in S phase in a concentration-dependent manner, and inducing apoptosis, whereas it had no significant inhibitory effects on the normal human IMR-90 fibroblasts. Furthermore, early and total induction of apoptosis was positively correlated with the concentration of CYBYT in SPC-A-1 cells, and the rate of total apoptosis was greater in the CYBYT 100 µg/mL and 50 µg/mL groups than that of the positive control 5-fluorouracil (5-Fu) group. Moreover, CYBYT upregulated bax, cleaved caspase-3 protein expression, downregulated bcl-2 protein expression, and released mitochondrial cytochrome c into the cytosol in a time- and concentration-dependent manner. Our findings indicated that CYBYT could significantly inhibit growth and induce apoptosis via the mitochondrial pathway in human lung adenocarcinoma cell line SPC-A-1.

High expression of serum miR-21 and tumor miR-200c associated with poor prognosis in patients with lung cancer.

Serum microRNAs have been identified as potential cancer biomarkers. However, the detailed mechanism by which expression of microRNAs contributes to the development and diagnosis of NSCLC remains unknown. This study was to identify specific miRNAs for diagnosing or predicting the prognosis of NSCLC patients and their correlation between miRNA expression in tissues and serums. Six matched cancer and noncancerous tissues from NSCLC patients were analyzed by miRNA microarray. Among these, three miRNAs (miR-21, miR-141, and miR-200c) were examined in 70 NSCLC paired samples (cancer, normal tissue, and serum) and 44 serum samples of normal volunteers by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Consisting with the microarray results, the expression levels of miR-21, miR-141, and miR-200c in NSCLC were higher than those in normal tissues. While the level of serum miR-21 was increased in cancer patients as compared with that in normal counterpart, expression of miR-141 and miR-200c showed lower levels in serums from cancer patients. Overexpression of serum miR-21 was strongly associated with lymph node metastasis and advanced clinical stage of NSCLC. Finally, log-rank and Cox regression tests demonstrated that high expressions of tumor miR 21 and miR-200c or serum miR-21 were associated with a poor survival in NSCLC patients. Our results suggest that tumor miR-21, miR-141, miR-200c, and serum miR-21 may be potential novel biomarkers for the diagnosis of NSCLC. In addition, this study, for the first time, identifies a significant role of the tumor miR-200c played in predicting prognosis in patients with NSCLC.

Prognostic evaluation of CapG, gelsolin, P-gp, GSTP1, and Topo-II proteins in non-small cell lung cancer.

Metastasis and multidrug resistance (MDR) are the main reasons for the poor prognosis of non-small cell lung cancer (NSCLC) patients. The use of biomarkers may contribute to a more accurate prediction of tumor metastasis, a better response to chemotherapy, and better patient survival. Gelsolin-like actin-capping protein (CapG) and gelsolin have been identified as playing important roles in tumor invasion and metastasis. Permeability glycoprotein (P-gp), glutathione S-transferase pi (GSTP1), and topoisomerase-II (Topo-II) are proteins that are closely related to MDR. In this study, we assessed the prognostic significance of CapG and gelsolin (both markers of tumor motility), and of P-gp, GSTP1, and Topo-II (markers of MDR) in NSCLC patients. One hundred and twenty-one patients with pathologically confirmed, resectable NSCLC were included in the study. The expression levels of the five kinds of proteins mentioned above were determined by immunohistochemistry (IHC). The correlation between the clinical characteristics and IHC findings were analyzed. Expression of CapG, gelsolin, and P-gp was found to be associated with an increased risk of death (Hazard Ratio (HR) = 2.799, 95% Confidence Interval (CI) = 1.2705-6.169, P = 0.011; HR = 3.968, 95% CI = 1.811-8.693, P = 0.001; HR = 3.251, 95% CI = 1.456-7.260, P = 0.004, respectively), whereas expression of GSTP1 and Topo-II was not. These results suggest that higher tumor motility and MDR may be important in NSCLC prognosis.

An integrated analysis of miRNA and mRNA expressions in non-small cell lung cancers.

Using DNA microarrays, we generated both mRNA and miRNA expression data from 6 non-small cell lung cancer (NSCLC) tissues and their matching normal control from adjacent tissues to identify potential miRNA markers for diagnostics. We demonstrated that hsa-miR-96 is significantly and consistently up-regulated in all 6 NSCLCs. We validated this result in an independent set of 35 paired tumors and their adjacent normal tissues, as well as their sera that are collected before surgical resection or chemotherapy, and the results suggested that hsa-miR-96 may play an important role in NSCLC development and has great potential to be used as a noninvasive marker for diagnosing NSCLC. We predicted potential miRNA target mRNAs based on different methods (TargetScan and miRanda). Further classification of miRNA regulated genes based on their relationship with miRNAs revealed that hsa-miR-96 and certain other miRNAs tend to down-regulate their target mRNAs in NSCLC development, which have expression levels permissive to direct interaction between miRNAs and their target mRNAs. In addition, we identified a significant correlation of miRNA regulation with genes coincide with high density of CpG islands, which suggests that miRNA may represent a primary regulatory mechanism governing basic cellular functions and cell differentiations, and such mechanism may be complementary to DNA methylation in repressing or activating gene expression.

[Promoter methylation status of hPer3 gene in AML patients and the in vitro effect of decitabine on the status].

OBJECTIVE: To investigate the clinical significance of promoter methylation status of hPer3 gene in acute myeloid leukemia (AML) patients and the in vitro effect of decitabine (DCA) on AML cell lines HL-60 and U937. METHODS: The promoter methylation status of hPer3 gene and mRNA expression levels in bone marrow of 206 AML and 40 iron deficiency anemia (IDA) patients (as control) were detected by methylation specific PCR (MS-PCR) and real-time PCR (RT-PCR). The HL-60 and U937 cell lines were treated with different concentrations of DCA for 48 and 72 h. The inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium (MTT); the early apoptosis rates by staining with Annexin V and PI; the CD14 and CD11b expressions by flow cytometry (FCM); the promoter methylation status of hPer3 gene by MS-PCR; and the hPer3 mRNA expressions levels by RT-PCR. RESULTS: The promoter methylation rates of hPer3 in newly diagnosed (ND) group, partial remission(PR) group, complete remission (CR) group, relapse (R) group and control group were 93.65% (59/63), 54.39% (31/57), 24.66% (18/73), 61.54% (8/13) and 0% (0/40), and the hPer3 mRNA expression levels were 0.19 ± 0.08, 6.28 ± 2.11, 52.76 ± 14.17, 8.18 ± 4.36, 75.03 ± 18.16, respectively. There was a significant statistic difference between any two group (P < 0.01) excepting for between PR and R group (P > 0.05). After DCA treatment, the promoter hypermethylation status of hPer3 was reduced and the mRNA and CD14, CD11b expression levels were up regulated in a dose dependent manner with an induction of cell apoptosis. CONCLUSIONS: Promotor methylation status and mRNA expression of hPer3 gene may be indicators for evaluating AML. DCA can induce the expression of hPer3 gene and cells apoptosis in AML.

Chan-Yu-Bao-Yuan-Tang induces apoptosis in NSCLC and SCLC cell lines via a mitochondria-mediated pathway.

Previous clinical studies have shown efficacy and safety of the traditional Chinese medicinal herb extract Chan-Yu-Bao-Yuan-Tang (CYBYT) in lung cancer patients. The effects of CYBYT on proliferation and apoptosis in the non-small cell lung cancer cell line NCI-H460 and the small cell lung cancer cell line NCI-H446 were investigated in vitro. CYBYT significantly induced antiproliferative effects of NCI-H460 and NCI-H446 cells in a concentration- and time-dependent manner. The IC(50) values in NCI-H460 cells were 94.37 µg/mL (24 h) and 20.89 µg/mL (48 h), whereas in NCI-H446 cells IC(50) values were 214.72 µg/mL (24 h) and 114.58 µg/mL (48 h). Annexin V-FITC/PI staining showed CYBYT could significantly induce apoptosis in NCI-H460 and NCI-H446 cells, and the total apoptosis rates were positively correlated with the concentration and time of CYBYT treatment. Furthermore, treatment with the broad-spectrum caspase inhibitor (z-VAD-fmk) effectively protected NCI-H460 and NCI-H446 cells against CYBYT-triggered apoptosis. The apoptotic processes involved were a marked decrease in antiapoptotic protein Bcl-2 and an increase in proapoptotic protein Bax. The release of mitochondrial cytochrome c into the cytosol was also observed, which, in turn, resulted in the activation of caspase-9 and caspase-3. CYBYT exerts antiproliferative and growth inhibition effects on NCI-H460 and NCI-H446 cells through the mitochondrial caspase-dependent cell death pathway.

[Expression and clinical significance of ZO-1 in patients with non-small cell lung cancer].

BACKGROUND AND OBJECTIVE: It has been proven that the expression of ZO-1 (zonula occluden-1) was correlated with the growth and metastasis of cancer. The aim of this study is to investigate the relationship between ZO-1 gene expression and clinicopathologic parameters and the survival rates in the patients with non-small cell lung cancer (NSCLC). METHODS: The expression levels of ZO-1 gene in 101 patients with NSCLC and lung tissues of 61 patients with benign lung disease were detected by real-time PCR, Western blot and immunohistochemical techniques. RESULTS: There was a significant statistic difference of ZO-1 mRNA expression levels between carcinoma group and control group (P < 0.01). In carcinoma group, adjacent group and control group, ZO-1 protein expression levels were with significantly statistic difference between each other (P < 0.01 or P < 0.05). There was a significantly statistic difference between ZO-1 mRNA expression levels in survival during 2 years follow-up group and death during 2 years follow-up group (t=-5.61, P < 0.01). CONCLUSIONS: ZO-1 is a valuable predictive marker for NSCLC patients.

[Expression and prognostic value of vacuole membrane protein 1 in non-small-cell lung cancer].

OBJECTIVE: To investigate the expression and prognostic value of vacuole membrane protein 1 (VMP1) in non-small-cell lung cancer (NSCLC). METHODS: The clinical data and survival status of 78 NSCLC patients were collected. Their paraffin-embedded tissues were detected immunohistochemically with a custom-made rabbit anti-human VMP1 polyclonal antibody. And the clinical significance of VMP1 expression and its relationship with the overall survival rate were analyzed statistically. RESULTS: The positive expression of VMP1 could only be observed in cytoplasm of cancer cells. Among 78 patients, 40 (51.3%) patients were stained VMP1-positive. The positive rate of VMP1 had a correlation with clinical stage (χ(2) = 6.829, P < 0.05), but not with gender, age, pathological type, gross classification and differentiation grade (P > 0.05). The overall 1-and 3-year survival rate was 94.9% and 66.3% respectively. The overall estimated survival time was 37.2 months (95%CI: 33.7 - 40.8). The expression of VMP1 had significant influence on the survival rate (χ(2) = 6.192, P < 0.05). The VMP1-positive patients lived shorter with an estimated survival time of 29.0 months (95%CI: 25.0 - 33.0). VMP1 expression and clinical stage were independent risk factors of influencing the survival rate. CONCLUSION: The detection of VMP1 in resected NSCLC tumor tissues may be helpful for prognostic prediction. NSCLC patients with VMP1-positive or late clinical stage have a worse prognosis.