SREBP1 site 1 protease inhibitor PF-429242 suppresses renal cell carcinoma cell growth.

Renal cell carcinoma (RCC) cells have increased lipogenesis and cholesterol synthesis. Sterol regulatory element-binding protein-1 (SREBP1) is cleaved by site 1 protease (S1P) to release the transcriptionally active amino-terminal domain. PF-429242 is a potent and competitive S1P inhibitor. We here tested its activity in RCC cells. In established and primary human RCC cells, PF-429242 potently inhibited cell proliferation, migration, and invasion. The S1P inhibitor provoked apoptosis activation in RCC cells. Furthermore, shRNA-mediated S1P silencing or CRISPR/Cas9-induced S1P knockout led to RCC cell growth inhibition and apoptosis activation. Conversely, ectopic overexpression of SREBP1 or S1P augmented RCC cell proliferation and migration. Daily i.v. injection of a single dose of PF-429242 robustly inhibited RCC xenograft growth in severe combined immunodeficiency mice. Additionally, intratumoral injection of S1P shRNA lentivirus inhibited RCC xenograft growth in mice. SREBP1, S1P, and its target gene low density lipoprotein receptor (LDLR) were significantly elevated in human RCC tissues. These results suggest that targeting S1P by PF-429242 inhibited RCC cell growth in vitro and in vivo.

Laboratory data analysis of novel coronavirus (COVID-19) screening in 2510 patients.

BACKGROUND: Novel coronavirus (COVID-19) is highly infectious and requires early detection, isolation, and treatment. We tried to find some useful information by analyzing the covid-19 screening data, so as to provide help for clinical practice. METHOD: We collected nucleic acid and hematology data from 2510 patients for COVID-19 infection for retrospective analysis. RESULT: COVID-19 and influenza A and B infection rates were 1.3%, 3%, and 3%, respectively. COVID-19 nucleic acid was detected in stool but not in tear samples from 8 positive patients. Among the 32 patients with COVID-19, 15 (47%) and 16 (50%) patients showed decreased lymphocyte count and lymphocyte ratio, 21(66%) and 24(75%) patients showed decreased eosinophil count and eosinophil ratio, and 18 (56%) patients showed increased C-reactive protein. Ten hematological indicators significantly differed in the blood of patients with COVID-19 and those with influenza A and B (P < 0.05). Eighteen hematological indicators significantly differed between patients with COVID-19 and negative patients (P < 0.05). CONCLUSION: The positive rate of influenza A and B infection was higher than that of COVID-19. When pharyngeal swab collection may cause infection, fecal samples can be examined. Evaluation of pharyngeal swab and fecal samples can improve the positive rate of nucleic acid detection. The COVID-19 can cause some hematological indices changes.

Long non-coding RNA expressed in macrophage co-varies with the inflammatory phenotype during macrophage development and polarization.

Advances in microarray, RNA-seq and omics techniques, thousands of long non-coding RNAs (lncRNAs) with unknown functions have been discovered. LncRNAs have presented a diverse perspective on gene regulation in diverse biological processes, especially in human immune response. Macrophages participate in the whole phase of immune inflammatory response. They are able to shape their phenotype and arouse extensive functional activation after receiving physiological and pathological stimuli. Emerging studies indicated that lncRNAs participated in the gene regulatory network during complex biological processes of macrophage, including macrophage-induced inflammatory responses. Here, we reviewed the existing knowledges of lncRNAs in the processes of macrophage development and polarization, and their roles in several different inflammatory diseases. Specifically, we focused on how lncRNAs function in macrophage, which might help to discover some potential therapeutic targets and diagnostic biomarkers.

The reference intervals for CA125, CA15-3, CA19-9, CA72-4, AFP, CEA, NSE and CYFRA21-1.

Tumor markers are noninvasive diagnostic tools for cancer. Their abnormal expression often occurs earlier than clinical symptoms or other detection signals. Appropriate reference intervals (RIs) of tumor markers are important for health evaluation, cancer diagnosis, therapy monitoring and prognosis assessment. In this study, we aimed to establish the RIs of cancer antigen 125 (CA125), CA15-3, CA19-9, CA72-4, alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), neuron-specific enolase (NSE) and cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) in apparently healthy Henan population. A total of 1705 apparently healthy participants (21-89 years) were recruited from five representative geographical regions in Henan province. Nonparametric 95th percentile intervals were used to define the RIs of CA125, CA15-3, CA19-9, CA72-4, AFP, CEA, NSE and CYFRA21-1. The test results of CA125, CA15-3, CA19-9, CA72-4, AFP, CEA, NSE and CYFRA21-1 can traceable to reference measurement procedures. The age- and gender-specific RIs of the tumor markers were established. We established age- and gender-specific RIs for CA125, CA15-3, CA19-9, CA72-4, AFP, CEA, NSE and CYFRA21-1. The newly established RIs should be more suitable for Henan population. It will be valuable for clinicians to make a medical diagnosis, therapeutic management decision and other physiological assessment.

Inhibition of constructed SEC3-ES lentiviral vector to proliferation, migration of Hela cells.

AIM: To construct a lentiviral vector with endostatin (ES) and staphylococcal enterotoxin C3(SEC3) gene, and investigate its capacities of inhibition on proliferation and migration of Hela cells. METHODS: By inserting ES and SEC3 gene into the plasmid and then transfect 293 T cell, the co-expressed (SEC3-ES) vector were constructed. A series of experiments in vitro were carried out to detect its anti-tumor capacity. RESULTS: SEC3 expression of the vector is about 3 times of GV365-SEC3 vector, and ES expression is over 22.5-fold compared with GV365-ES vector. Moreover, OD490 value of CO group (1.212 ± 0.003) was notably lower than NC (negative control) group (1.124 ± 0.01) (P < 0.05) in MTT assay. Cell cycle analysis showed it could block Hela cells in S phase. Meanwhile, in wound healing assay, cells of CO group migrated at a slower rate (0.59 ± 0.02) compared with NC group (0.65 ± 0.02)(P < 0.01). CONCLUSION: The successful construction of co-expressed vector lays the foundation for further studies in vivo. These promising results suggest a new strategy to treating cervical cancer.

The role of lncRNAs in signaling pathway implicated in CC.

Cervical cancer (CC) is the second most common malignancy in females. Owing to poor diagnosis, resistance to the systemic therapies, and high recurrence rate, patients with CC have a relatively poor prognosis. The role of a signaling pathway in CC has always been the focus among worldwide researchers. As reported before, aberrant expression of proteins associated with signaling pathways, such as phosphatidylinositol 3-kinase(PI3K), EGF-R, ß-catenin, and Erk and Bcl-2 was discovered in CC. Therefore, aberrant molecular signaling pathways are significant parts of cervical carcinogenesis. Recently discovered long noncoding RNAs (lncRNAs) as a new regulator player of molecular biology in CC have always been reported. In this review, we highlighted the role of lncRNA in signaling pathway implicated in CC and outlined the molecular mechanism of lncRNA in it. All of these present an opportunity for developing diagnosis and therapies against CC.

Assessment of a combination of Serum Proteins as potential biomarkers to clinically predict Schizophrenia.

Schizophrenia (SZ) is a devastating psychiatric disorder. Validation of potential serum biomarkers during first-episode psychosis (FEP) is especially helpful to understand the onset and prognosis of this disorder. To address this question, we examined multiple blood biomarkers and assessed the efficacy to diagnose SZ. The expression levels of Neuregulin1 (NRG1), ErbB4, brain-derived neurotrophic factor (BDNF), DNA methyltransferases 1 (DNMT1) and ten-eleven translocation 1 (TET1) proteins in peripheral blood of 53 FEP patients and 57 healthy controls were determined by enzyme-linked immunosorbent assay (ELISA). Multivariable logistic regression including biomarker concentration as covariates was used to predict SZ. Differentiating performance of these five serum protein levels was analyzed by Receiver Operating Characteristic (ROC) curve analysis. We found that patients with SZ present a higher concentration of DNMT1, and TET1 in peripheral blood, but a lower concentration of NRG1, ErbB4 and BDNF than controls. Multivariable logistic regression showed that ErbB4, BDNF and TET1 were independent predictors of SZ, and when combined, provided high diagnostic accuracy for SZ. Together, our findings highlight that altered expression of NRG1, ErbB4, BDNF, DNMT1 and TET1 are involved in schizophrenia development and they may serve as potential biomarkers for the diagnosis of the schizophrenia. Therefore, our study provides evidence that combination of ErbB4, BDNF and TET1 biomarkers could greatly improve the diagnostic performance.

Wnt signaling in cervical cancer?

Cervical cancer (CC) is the second most common malignant cancer in women. CC is difficult to diagnose, has a high recurrence rate, and is resistant to systemic therapies; as a result, CC patients have a relatively poor prognosis. One potential link to CC is the Wnt signaling pathway and its downstream effectors, which regulate cell differentiation, proliferation, migration, and fate. The aberrant activation of Wnt signaling is associated with various cancers, including CC. Recent studies have shown that activating or inhibiting the intracellular signal transduction in this pathway can regulate cancer cell growth and viability. This review will summarize the experimental evidence supporting the significance of the Wnt signaling pathway in CC, and will also discuss the current clinical role of Wnt signaling in CC diagnosis, therapy, and prognosis.

In vitro effects of Staphylococcus aureus enterotoxin C3 on T cell activation, proliferation and cytokine production.

The present study aimed to investigate the effects of Staphylococcus aureus enterotoxin C3 (SEC3), including recombinant (r)SEC3 protein and lentivirus­mediated SEC3, on the activation, proliferation and cytokine production of human T cells. HeLa cells were infected with SEC3 lentiviral vector (LV­SEC3) and viability was determined using the Cell Counting Kit­8 (CCK­8) assay. Subsequently, infected cells or rSEC3 protein were co­cultured with human peripheral blood mononuclear cells (PBMCs) for 10 days, after which the culture supernatant and T cells were incubated with untreated HeLa cells, which were subjected to a CCK­8 assay to determine cytotoxicity. In addition, IL­6 and IFN­Î³ expression was detected by chemiluminescence and enzyme­linked immunospot analyses, respectively. Subpopulations of activated T cells were sorted by flow cytometry. The results demonstrated that, following infection with LV­SEC3 or negative control lentiviral vector (LV­NC), >80% of HeLa cells presented green fluorescent protein­positive signals. All five groups of co­cultured T cells exhibited proliferation. Co­culture of PBMCs with rSEC3 protein or LV­SEC­infected cells resulted in elevated IL­6 and IFN­Î³ secretion. In addition, rSEC3­activated and monocultured T cells were predominantly cluster of differentiation (CD)4+ (62.7 and 59.6%, respectively) whereas phytohemagglutinin­stimulated T cells were predominantly CD8+ (57.8%). Compared with the LV­NC group, T cells and culture supernatants from the LV­SEC3 group significantly attenuated proliferation of HeLa cells. These results suggest that rSEC3 protein, and LV­SEC3­infected HeLa cells, are able to potently activate T cells, increasing cytokine production and amplify the antitumor immune response.

The age-specific reference intervals for tPSA, fPSA, and %fPSA in healthy Han ethnic male.

INTRODUCTION: Prostate cancer (PC) is one of the most common malignancies in male, and has become the fastest growing malignancy in recent years. Prostate specific antigen (PSA) is widely used as a tumor marker to screen for PC. Many studies have been performed to define the reference intervals (RIs) for total circulating PSA (tPSA). The results were different among different nations and races, even in the same race. Few researches have been performed on the RIs of free PSA (fPSA) and the ratio of free to total PSA (%fPSA). In this study, we aimed to simultaneously determine the age-specific RIs for tPSA, fPSA, and %fPSA in the healthy Han ethnic male. METHODS: A total of 1862 apparently healthy male aged from 21 to 94 years were included in our study. Nonparametric 95th percentile intervals were used to define the RIs. RESULTS: The reference limits in different age groups (21-50, 51-60, 61-70, 71-80, and ≥81 years) were 2.07, 3.59, 4.93, 6.83, and 7.73 ng/mL for tPSA, and 0.60, 0.76, 0.83, 1.30, and 2.41 ng/mL for fPSA. The RIs of %fPSA were ≥0.16 for 21-50 years and ≥0.13 for male over 50 years old. CONCLUSIONS: We established age-specific RIs for tPSA, fPSA and %fPSA. The newly established RIs should be more suitable for Chinese Han ethnic male. It will be valuable for physicians to make exact medical decision and appropriate medical intervention.

CHN2 Promoter Methylation Change May Be Associated With Methamphetamine Dependence.

BACKGROUND: Methamphetamine (MA) abuse is becoming increasingly serious in China. The mechanism of MA dependence remains unclear. CHN2 gene encodes chimeric protein-2 that regulate axonal pruning via the Rac-GTPase system and play a pivotal role in the formation of nervous circuits. Genetic studies suggest that the polymorphism of the CHN2 gene was related to substance dependence. AIMS: The aim of this study was to investigate the association between the methylation of CHN2 gene promoter with MA dependence. METHODS: According to SCID-I (Structured Clinical Interview for DSM-IV Axis I Disorders, SCID-I) used for investigating MA dependence, 224 male MA addicts were recruited into the case group. In addition, 109 healthy men were recruited into the control group. Blood samples were collected with the purpose of detecting the methylation levels of CHN2 gene promoter by methylight qPCR. The association between the methylation of CHN2 gene promoter with MA dependence was analyzed. RESULTS: The mean (sd) methylation levels of CHN2 gene promoter in the case group were significantly higher than in the control group, which were 2795.55 (733.19) and 1026.73 (698.73), respectively, showing significant differences between the two groups (t=21.25, p<0.001). Pearson analysis showed no significant correlation between the methylation levels of CHN2 promoter and other factors (the age of initial MA use, the duration of MA use, combination with K powder, tobacco and alcohol). CONCLUSIONS: The abnormal methylation of CHN2 gene promoter was significantly correlated with MA dependence.

Sub-chronic Antipsychotic Drug Administration Reverses the Expression of Neuregulin 1 and ErbB4 in a Cultured MK801-Induced Mouse Primary Hippocampal Neuron or a Neurodevelopmental Schizophrenia Model.

It has been reported that specific environmental influences during the postpartum period might contribute to the development of schizophrenia (SZ). Administration of MK801 during early development led to persistent brain pathology. Glutamate decarboxylase 1 (GAD67) and parvalbumin (PV), and neuregulin 1 (NRG1)/ErbB4 signaling were closely associated with SZ pathology. We postulated therefore that NMDA receptor antagonists exposure during the postpartum period may be associated with expression dysregulation of some of the SZ candidate proteins. To test this, we used mouse primary hippocampal neurons and neonatal male mice treated with the NMDA receptor antagonist, MK801 at postnatal day 4 (P4) or P7, followed by the treatments of antipsychotic drugs (i.e., olanzapine, risperidone, and haloperidol). The expressions of GAD67, PV, NRG1, and ErbB4 in in vitro and in vivo SZ models were detected with Western blot analysis and immunohistochemistry, respectively. Behavioral tests (locomotion activity, social interaction, novel object recognition and prepulse inhibition) were measured. We found MK801 decreased the expression of GAD67, PV, NRG1 and ErbB4, and induced obvious behavioral alterations, while antipsychotics reversed these alterations. These results suggest that exposure to the NMDA receptor antagonist in early development may lead to long-lasting influence on the expression of specific proteins, such as GAD67, PV, NRG1, and ErbB4. Moreover, our results suggest that rescue of the activation of the NRG1/ErbB4 signaling pathway may be one of the mechanisms by which antipsychotic drugs have an antipsychotic effect.

Serum pepsinogen reference intervals in apparently healthy Chinese population with latex enhanced turbidimetric immunoassay.

AIM: Serum pepsinogen (sPG) has been used to help in diagnosing atrophic corpus gastritis and in screening for gastric cancer non-invasively. There are as yet no reports on sPG reference intervals (RIs) with latex enhanced turbidimetric immunoassay (LIA). In this study, we established the RIs for sPG in a healthy Chinese population using LIA. METHODS: Serum PGI and PGII levels in a healthy population (aged 17-80 years) were measured simultaneously using LIA. RIs were determined following Clinical Laboratory and Standards Institute C28-A3 guidelines using a non-parametric method. RESULTS: 95% RIs in men (ng/mL) were: ≤40 years old, 25.53-100.76 for PGI and ≤24.42 for PGII; 41-50 years old, 26.62-124.74 for PGI and ≤26.81 for PGII; and 51-80 years old, 30.40-153.25 for PGI and ≤32.62 for PGII. Corresponding RIs for women (ng/mL) were: ≤40 years old, 21.20-87.44 for PGI and ≤25.53 for PGII; and 41-80 years, 26.40-127.46 for PGI and ≤30.18 for PGII. 95% RI for PGI/PGII in both men and women at any age was ≥2.51. CONCLUSIONS: We established the RIs for sPG using LIA in a healthy Chinese population, which can provide a reference for clinical and laboratory studies.

The reference intervals for HE4, CA125 and ROMA in healthy female with electrochemiluminescence immunoassay.

OBJECTIVES: Human epididymis protein 4 (HE4) is a tumor marker in diagnosing ovarian cancer. Together with cancer antigen 125 (CA125) and the risk of ovarian malignancy algorithm (ROMA) score, it can improve sensitivity and specificity. There are no reports about the serum HE4 reference intervals (RIs) with electrochemiluminescence immunoassay (ECLIA). The RI of ROMA hasn't been established yet. In this study, we establish the RIs for serum HE4, CA125 and ROMA in healthy Chinese female with ECLIA. METHODS: Serum HE4 and CA125 concentrations in healthy female (age ranged from 21 to 81 years) were simultaneously measured with ECLIA on Roche Cobas E601 system. The RIs were determined following CLSI C28-A3 guidelines using a nonparametric method. RESULTS: The upper limits of the 95% percentile intervals were 82.62 pmol/L for HE4, 30.91 U/mL for CA125 and 19.27 for ROMA. The reference limits for HE4, CA125 and ROMA were 65.87 pmol/L, 32.23 U/mL, 13.14 for premenopausal women, and 90.76 pmol/L, 27.52 U/mL, 25.46 for postmenopausal women respectively. CONCLUSION: We established the RIs for serum HE4, CA125 and ROMA with ECLIA in healthy Chinese female. It provided a reference for clinical and laboratory studies.

Aberrant expression of E-cadherin in lung tissues of patients with probable lung cancer.

INTRODUCTION: This study assessed the relationship of E-cadherin mRNA and protein expression with the diagnosis of lung cancer with the aim of providing an auxiliary diagnostic method. METHODS: Semi-quantitative nested RT-PCR and western blotting were applied to detect E-cadherin mRNA transcripts and protein, respectively, in 30 cases of diagnostic lung cancer, 30 cases of clinically suspected patients with lung cancer and 30 cases of other disease. Immunohistochemical staining was also used to detect E-cadherin. RESULTS: Remarkably decreased levels of relative E-cadherin mRNA value and increased E-cadherin protein negativity were observed in probable lung cancer, when compared with possible lung cancer and others. With a threshold of 1.45, relative E-cadherin mRNA value showed a sensitivity of 90% and a specificity of 83% for the diagnosis of lung cancer. The combination of decreased relative E-cadherin mRNA value and negative E-cadherin protein increased the specificity and sensitivity. CONCLUSION: These data suggest that Chinese patients with diagnostic lung cancer have similar decreased levels of relative E-cadherin mRNA and E-cadherin protein value in the lung cancer tissues as in lung cancer patients in other countries. Measurement of relative E-cadherin mRNA and protein values in lung cancer tissues has potential for lung cancer diagnosis.

Clinical significance of simultaneous determination of serum tryptophan and tyrosine in patients with lung cancer.

BACKGROUND: To explore the clinical significance of serum tyrosine (Tyr) and tryptophan (Trp) in patients with lung cancer, we used a simple and efficient method of high-performance liquid chromatography with fluorescence detection (HPLC-FD) that simultaneously measured serum Trp and Tyr contents. METHODS: The concentrations of Tyr and Trp were measured simultaneously by HPLC-FD in the sera of 80 patients with lung cancer and 120 healthy controls. RESULTS: Trp concentrations were significantly lower in patients with lung cancer than in healthy controls (39.26±5.44 vs. 49.93±5.43 µmol/l, respectively; P<0.01), whereas in Tyr concentrations there were no differences with healthy controls (65.38±7.94 vs.66.40±8.55 µmol/l, respectively; P>0.05). In addition, patients in the adenocarcinoma group had significantly lower Trp and Tyr concentrations than those in squamous cell carcinoma group. There was no difference between the early stage and advanced stage of lung cancer. CONCLUSIONS: Determination of serum Trp and Tyr concentrations can be employed to assist the diagnosis of the histotypes of lung cancer and tumor stage. Tyr and Trp as indexes on the lung cancer diagnostic sensitivity, specificity were 54.9, 62.9% and 82.4, 92.1%, Trp is an important and special index for lung cancer diagnosis of which the specificity of diagnosis of lung cancer is more than 92%.

Insulin receptor substrate 1 regulates the cellular differentiation and the matrix metallopeptidase expression of preosteoblastic cells.

Insulin receptor substrate 1 (IRS1) is an essential molecule for the intracellular signaling of IGF1 and insulin, which are potent anabolic regulators of bone metabolism. Osteoblastic IRS1 is essential for maintaining bone turnover; however, the mechanism underlying this regulation remains unclear. To clarify the role of IRS1 in bone metabolism, we employed RNA interference to inhibit IRS1 gene expression and observed the effects of silencing this gene on the proliferation and differentiation of and the expression of matrix metallopeptidase (MMP) and tumor necrosis factor receptor superfamily, member 11b (TNFRSF11B) in MC3T3-E1 cells. Our results showed that IRS1 short hairpin RNAs can effectively suppress the expression of IRS1, and inhibit the phosphorylation of AKT in IRS1 pathway; reduce the expression of MMP2, MMP3, MMP13, and MMP14, decrease the expression of TNFRSF11B and RANKL (also known as tumor necrosis factor (ligand) superfamily, member 11), however increase the RANKL/TNFRSF11B ratio; decrease cell survival, proliferation, and mineralization, and impair the differentiation of MC3T3-E1 cells. The downregulation of IRS1 had no effect on the expression of MMP1. Our findings suggest that IRS1 not only promotes bone formation and mineralization but also might play roles in bone resorption partly via the regulation of MMPs and RANKL/TNFRSF11B ratio, thus regulates the bone turnover.