Diagnostic and prognostic value of serum soluble B7-H3 in nonsmall cell lung cancer.

The aim of this study was to investigate the utility of serum soluble B7-H3 (sB7-H3) as a diagnostic marker for early-stage nonsmall cell lung cancer (NSCLC) and its potential for evaluating the prognosis of patients with advanced-stage NSCLC. In this study, an ELISA was employed to detect the expression levels of sB7-H3 in a cohort of patients diagnosed with NSCLC ( n = 122) and a control group ( n = 42) during the same observation period. Comparative analyses were conducted to ascertain the variations in sB7-H3 concentrations between the NSCLC cohort and the healthy control group, as well as across pathological types and the presence and absence of lymph node metastasis. (1) The concentration of sB7-H3 in patients diagnosed with NSCLC exhibited a statistically significant increase compared to that observed in the healthy control group ( P < 0.05). Elevated expression levels of sB7-H3 demonstrated a significant correlation with pathological type, lymph node metastasis, tumor, node and metastasis stage and programmed cell death ligand (PD-L1) expression ( P < 0.05). (2) The diagnostic utility of sB7-H3 for the diagnosis of NSCLC and the heightened expression of PD-L1 demonstrated high levels of sensitivity and specificity. (3) Elevated levels of sB7-H3 emerged as an independent risk factor impacting the overall survival of patients diagnosed with advanced NSCLC. The findings of this study suggest that sB7-H3 holds promise as a diagnostic tool for early-stage NSCLC. The elevated expression of sB7-H3 appears to serve as a reliable indicator for assessing the prognosis of patients diagnosed with advanced NSCLC.

Pesticin-Like Effector VgrG3cp Targeting Peptidoglycan Delivered by the Type VI Secretion System Contributes to Vibrio cholerae Interbacterial Competition.

Vibrio cholerae can utilize a type VI secretion system (T6SS) to increase its intra- and interspecies competition. However, much still remains to be understood about the underlying mechanism of this intraspecies competition. In this study, we isolated an environmental V. cholerae strain E1 that lacked the typical virulence factors toxin-coregulated pilus and cholera toxin and that encoded a functional T6SS. We identified an evolved VgrG3 variant with a predicted C-terminal pesticin-like domain in V. cholerae E1, designated VgrG3cp. Using heterologous expression, protein secretion, and peptidoglycan-degrading assays, we demonstrated that VgrG3cp is a T6SS-dependent effector harboring cell wall muramidase activity and that its toxicity can be neutralized by cognate immunity protein TsiV3cp. Site-directed mutagenesis proved that the aspartic acid residue at position 867 is crucial for VgrG3cp-mediated antibacterial activity. Bioinformatic analysis showed that genes encoding VgrG3cp-like homologs are distributed in Vibrio species, are linked with T6SS structural genes and auxiliary genes, and the vgrG3cp-tsiV3cp gene pair of V. cholerae probably evolved from Vibrio anguillarum and Vibrio fluvialis via homologous recombination. Through a time-lapse microscopy assay, we directly determined that cells accumulating VgrG3cp disrupted bacterial division, while the cells continued to increase in size until the loss of membrane potential and cell wall breakage and finally burst. The results of the competitive killing assay showed that VgrG3cp contributes to V. cholerae interspecies competition. Collectively, our study revealed a novel T6SS E-I pair representing a new T6SS toxin family which allows V. cholerae to gain dominance within polymicrobial communities by T6SS. IMPORTANCE The type VI secretion system used by a broad range of Gram-negative bacteria delivers toxic proteins to target adjacent eukaryotic and prokaryotic cells. Diversification of effector proteins determines the complex bacterium-bacterium interactions and impacts the health of hosts and environmental ecosystems in which bacteria reside. This work uncovered an evolved valine-glycine repeat protein G3, carrying a C-terminal pesticin-like domain (VgrG3cp), which has been suggested to harbor cell wall hydrolase activity and is able to affect cell division and the integrity of cell wall structure. Pesticin-like homologs constitute a family of T6SS-associated effectors targeting bacterial peptidoglycan which are distributed in Vibrio species, and genetic loci of them are linked with T6SS structural genes and auxiliary genes. T6SS-delivered VgrG3cp mediated broad-spectrum antibacterial activity for several microorganisms tested, indicating that VgrG3cp-mediated antimicrobial activity is capable of conferring bacteria a competitive advantage over competitors in the same niches.

Application of three-dimensional Raman imaging to determination of the relationship between cellular localization of diesel exhaust particles and the toxicity.

A diesel exhaust particle (DEP) is a type of particulate matter that is easily produced from combustion in a diesel power engine. It has been reported that DEPs can cause short- and long-term health problems. This is because DEPs are complex mixtures that are highly inhalable through the airways due to their small particle size. However, the relationship between intracellular localization of DEPs after their deposition in the lungs and the subsequent biological responses remains to be clarified. This is due to difficulties in distinguishing particles that are inside the cells from those that are outside. In this study, A549 human lung epithelial cells were exposed to DEPs at concentrations of 0, 25, 75, or 200 µg/mL for different periods, after that particles in the A549 cells were analyzed by three-dimensional (3D) images obtained from a Raman microscope. The cytotoxic effects of DEPs on the A549 cells were investigated by measuring cell viability, the levels of intracellular reactive oxygen species (ROS) and cell death. The Raman microscopy revealed that the particles invaded the A549 cells, and at a concentration of 200 µg/mL, they markedly decreased cell viability, increased intracellular ROS production, triggered late apoptosis/necrosis and induced nuclear damage. These results suggest that intracellular DEPs exposed at a high concentration may be highly toxic and can impair the viability of A549 cells. Furthermore, the 3D images from the Raman microscopy can be used to evaluate intracellular particle dynamics.

Interleukin-18 promotes the antitumor ability of natural killer cells in colorectal cancer via the miR-574-3p/TGF-ß1 axis.

Interleukin (IL)-18 has a clear antitumor effect; however, its mechanisms of action are not understood in patients with colorectal cancer (CRC). Here, we investigated the potential mechanism of IL-18 in CRC. The results showed that IL-18 treatment alone had no effect on HCT116 cells apoptosis, whereas IL-18 in the presence of natural killer (NK) cells resulted in apoptosis and inhibition of cells proliferation in vitro. Profiling of miRNA expression following coculture with NK cells and treatment with IL-18 resulted in significant downregulation of miR-574-3p expression and upregulated expression of the target gene transforming growth factor beta 1 (TGF-ß1). miR-574-3p binds to TGF-ß1, and miR-574-3p overexpression increased the proliferation and decreased the apoptotic rate of HCT116 cells in NK cells coculture with IL-18 treatment; overexpression of TGF-ß1 restored the effect of miR-574-3p overexpression. The miRNA profile of HCT116 undergoes significant alteration before and after coculturing with NK cells and treatment with IL-18. IL-18 alone did not affect HCT116 cells apoptosis but did promote the antitumor ability of NK cells in coculture with HCT116 cells via the miR-574-3p/TGF-ß1 axis. Our study suggested that IL-18 can be a new potential target for cancer immunotherapy for CRC.

High prevalence and risk factors of multiple antibiotic resistance in patients who fail first-line Helicobacter pylori therapy in southern China: a municipality-wide, multicentre, prospective cohort study.

BACKGROUND: We aimed to study the prevalence of secondary antibiotic resistance of Helicobacter pylori in southern China and its risk factors, particularly geographical and socio-economic factors. METHODS: This was a municipality-wide, multicentre, prospective cohort study involving five major hospitals. Patients aged ≥18 years who failed first-line bismuth-based quadruple anti-H. pylori therapy between September 2016 and February 2018 were recruited. Participants underwent upper gastrointestinal endoscopy with biopsy from the antrum and body for H. pylori culture and antimicrobial susceptibility testing for six antibiotics (clarithromycin, levofloxacin, metronidazole, amoxicillin, tetracycline and furazolidone). Patients with failure of H. pylori culture were excluded. Participants completed a questionnaire profiling 22 potential risk factors of H. pylori infection and antibiotic resistance, including medical, social, household and birthplace factors. RESULTS: A total of 1113 patients failed first-line therapy, with successful H. pylori culture in 791 (71.1%) [male = 433 (54.7%); median age = 43 years]. Secondary resistance rates of dual antibiotics (clarithromycin + metronidazole and levofloxacin + metronidazole) and triple antibiotics (clarithromycin + levofloxacin + metronidazole) were 34.0%, 38.7% and 17.8%, respectively. Risk factors for clarithromycin + metronidazole resistance were history of ≥2 courses of H. pylori therapies [adjusted OR (aOR) = 1.71; 95% CI = 1.17-2.54], ≥3 household members (aOR = 2.00; 95% CI = 1.07-3.90) and family history of gastric cancer (aOR = 1.85; 95% CI = 1.18-2.85). Risk factors for levofloxacin + metronidazole resistance were age ≥40 years (aOR = 1.94; 95% CI = 1.37-2.75), lower gross domestic product per capita (aOR = 0.29; 95% CI = 0.10-0.80) and higher number of doctors/10 000 population (aOR = 1.59; 95% CI = 1.07-2.39). A higher human development index was of borderline significance (aOR = 2.79; 95% CI = 0.97-8.70). CONCLUSIONS: The rates of secondary resistance of H. pylori to multiple antibiotics were high in southern China. Certain population-level risk factors were associated with levofloxacin + metronidazole resistance.

Effect of PD98059 on chemotherapy in patients with colorectal cancer through ERK1/2 pathway.

PURPOSE: To investigate the effects of MAPK/ERK signaling transduction pathway inhibitor PD98059 combined with paclitaxel on the proliferative, invasive and apoptotic abilities of colorectal cancer cells. METHODS: Colorectal cancer (CRC) SW-480 cells were selected and divided into 3 groups after passage: Blank group (not treated), control group (treated with PD98059 blocker (25µmol/L)), observation group (treated with PD98059 blocker (25µmol/L) combined with paclitaxel (100µmol/L)). RESULTS: The proliferation, invasion and apoptosis of cells in each group were registered. The relative expressions of ERK1/2 and p-ERK1/2 protein were assessed by Western blot. The relative expressions of Bax and Bcl-2 were assessed by RT-PCR. The proliferation ability in the control group was significantly higher than that in the observation group (p<0.05). The relative expression of p-ERK1/2 protein in the control group was significantly higher than that in the observation group (p<0.05). The invasive ability in the blank group was significantly higher than that in the other two groups (p<0.05), and that in the control group was significantly higher than that in the observation group (p<0.05). The apoptotic ability in the control group was significantly lower than that in the observation group p<0.05). The expression of Bcl-2 in the cells of the observation group was significantly higher than that of the blank group and the control group, and that in the control group was higher than that in the blank group (p<0.05). The expression of Bax in the cells of the observation group was significantly lower than that of the blank group and the control group (p<0.05). CONCLUSIONS: Inhibitor PD98059 combined with paclitaxel can affect the expression of ERK1/2 in ERK1/2 signaling pathway, effectively inhibit the proliferation and invasive abilities of CRC cells, increase the apoptotic ability of CRC cells, and is expected to become a potential drug for clinical treatment of CRC.

The expressions and significance of B7-H3 and CTLA-4 in the clinical stages of non-small-cell lung cancer.

The aim of this study was to analyze the expressions and significance of B7-H3 and CTLA-4 in the clinical stages of non-small-cell lung cancer (NSCLC). Seventy patients with NSCLC who underwent surgical resection or biopsy between January 2016 and February 2018 were enrolled. Among them, 30 were cases of paracancerous tissues and were assigned to the control group (CON). These cases were analyzed using immunochemical methods. Of the 70 cases, 48 were of adenocarcinoma, 19 were of squamous cell carcinoma, and 3 were of adenosquamous carcinoma. The expression rates of B7-H3 and CTLA-4 in the observation group (OBS) were 64.2% and 57.1% respectively, and those in the CON group were 6.7% and 0%, respectively (χ2=27.988, 28.571, P<0.001). The expression levels of B7-H3 and CTLA-4 in patients with poor differentiation, in stages III-IV, or with lymph node metastasis were significantly higher than those in patients with good-to-moderate differentiation, in stages I-II, and without lymph node metastasis (P<0.05). There was a positive correlation between the expressions of B7-H3 and CTLA-4 in the OBS group (r=0.74, P<0.05). B7-H3 and CTLA-4 are highly expressed and positively correlated with each other in NSCLC patients and are also closely related to clinical stages.

Paclitaxel alleviated liver injury of septic mice by alleviating inflammatory response via microRNA-27a/TAB3/NF-κB signaling pathway.

Excessive inflammatory response and apoptosis play an important role in the sepsis-induced liver injury. Paclitaxel, a diterpene alkaloid of Taxus brevifolia, is widely used as an anti-tumor drug and shows protective effects on acute lung and kidney injury. However, whether it has a protective effect against sepsis-induced liver injury has not been reported. The objective of this study was to investigate the protective effects of paclitaxel in septic liver injury in mice and associated molecular mechanisms. Our results showed that paclitaxel treatment improved LPS-induced liver injury, as evidenced by the reduced aminotransferase activity, histological scores and apoptosis in the liver tissues. This was accompanied by the alleviating of inflammation and oxidative stress, such as decreased levels of tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-6) interleukin-1ß (IL-1ß) and malondialdehyde (MDA) and increased levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in serum and liver tissues. Subsequent microarray and qRT-PCR analysis further showed that miR-27a was significantly decreased in mice with sepsis, which was recovered by paclitaxel pretreatment. Antagomir-miR-27a suppressed the therapeutic effects of paclitaxel in mice liver injury model via promoting inflammatory response. Of note, TAB3, which participated in the activation of the NF-κB signaling pathway, was identified as a direct target of miR-27 by luciferase reporter gene assays. Then, we revealed a reverse relationship between miR-27a expression levels and TAB3 mRNA levels in liver tissues from septic mice. Furthermore, paclitaxel treatment significantly decreased the expression of NF-κB p65, but increased inhibitor of nuclear factor-κB-α (IκBα) protein levels in septic mice, suggesting the inactivation of NF-κB signaling pathway. Notably, the inhibitory effects of paclitaxel on NF-κB signaling pathway were reversed by antagomir-miR-27a. Our data indicated that paclitaxel significantly attenuated septic induced liver injury through reducing inflammatory response via miR-27a/TAB3/NF-κB signaling pathway.

Correction: MiR-34a-5p promotes the multi-drug resistance of osteosarcoma by targeting the CD117 gene.

[This corrects the article DOI: 10.18632/oncotarget.8546.].

The miR-34a-5p promotes the multi-chemoresistance of osteosarcoma via repression of the AGTR1 gene.

BACKGROUND: Chemoresistance hinders the curative cancer chemotherapy. MicroRNAs (miRNAs) are key players in diverse biological processes including the chemoresistance of cancers. METHODS: A RNA-seq-based miR-omic analysis of osteosarcoma (OS) cells was performed to detect the levels of miR-34a-5p. Bioinformatics analysis revealed that AGTR1 is one of the target genes of miR-34a-5p. The mRNA and protein levels of AGTR1 were detected in both the miR-34a-5p-mimic transfected G-292 and miR-34a-5p-antagomiR transfected SJSA-1 cells. The involvement of AGTR1 with OS chemoresistance was validated by the experiments with siRNA-mediated repression or overexpression of the AGTR1 gene. RESULTS: We showed that miR-34a-5p promotes the multi- chemoresistance of OS. The angiotensin II type 1 receptor (AGTR1) gene, is one of the targets of miR-34a-5p in OS and thus negatively correlates with OS chemoresistance by systematic investigations of a multi-drug sensitive (G-292) and resistant (SJSA-1) OS cell lines. Down-regulation of the AGTR1 expression by siRNA passivates G-292 cells and suppresses cell apoptosis, while over-expression of AGTR1 sensitizes SJSA-1 cells and thus promotes the drug-triggered cell death. CONCLUSIONS: The miR-34a-5p and its target gene AGTR1 are the potential targets for an effective chemotherapy of OS. Our results also provide novel insights into the effective chemotherapy for OS patients.

MiR-34a-5p promotes the multi-drug resistance of osteosarcoma by targeting the CD117 gene.

An association has been reported between miR-34a-5p and several types of cancer. Specifically, in this study, using systematic observations of multi-drug sensitive (G-292 and MG63.2) and resistant (SJSA-1 and MNNG/HOS) osteosarcoma (OS) cell lines, we showed that miR-34a-5p promotes the multi-drug resistance of OS through the receptor tyrosine kinase CD117, a direct target of miR-34a-5p. Consistently, the siRNA-mediated repression of CD117 in G-292 and MG63.2 cells led to a similar phenotype that exhibited all of the miR-34a-5p mimic-triggered changes. In addition, the activity of the MEF2 signaling pathway was drastically altered by the forced changes in the miR-34a-5p or CD117 level in OS cells. Furthermore, si-CD117 suppressed the enhanced colony and sphere formation, which is in agreement with the characteristics of a cancer stem marker. Taken together, our data established CD117 as a direct target of miR-34-5p and demonstrated that this regulation interferes with several CD117-mediated effects on OS cells. In addition to providing new mechanistic insights, our results will provide an approach for diagnosing and chemotherapeutically treating OS.

Clinical relevance of using autofluorescence bronchoscopy and white light bronchoscopy in different types of airway lesions.

OBJECTIVES: The aim of this study was to evaluate the sensitivity and specificity of autofluorescence bronchoscopy (AFB) according to the macroscopic appearance of airway lesions under white light bronchoscopy (WLB). MATERIALS AND METHODS: The bronchoscopic findings of 708 patients, who were pathologically and clinically diagnosed with airway lesions and underwent both WLB and AFB, were analyzed. RESULTS: We recruited 708 patients for this study, of which 336 (47.5%) had benign lesions; 300 and 254 benign lesions were detected by AFB (specificity, 89.3%) and WLB (specificity, 75.6%), respectively. In 372 (52.5%) patients with bronchiogenic carcinoma, 356 and 235 lesions were identified by AFB (sensitivity, 95.7%) and WLB (sensitivity, 63.2%), respectively. The sensitivity and specificity of AFB for diagnosing lung cancer were higher than those of WLB (P < 0.05). Moreover, AFB showed high sensitivity for detecting lung cancer in cases in which WLB revealed hyperplasia, infiltration, and stenosis (P < 0.05). CONCLUSIONS: AFB combined with WLB could effectively improve the diagnosis of airway lesions.

MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR, respectively.

MicroRNAs have been identified as key players in the development and progression of osteosarcoma, which is the most common primary malignancy of bone. Sequencing-based miR-omic and quantitative real-time PCR analyses suggested that the expression of miR-193a-3p and miR-193a-5p was decreased by DNA methylation at their promoter region in a highly metastatic osteosarcoma cell line (MG63.2) relative to their expression in the less metastatic MG63 cell line. Further wound-healing and invasion assays demonstrated that both miR-193a-3p and miR-193a-5p suppressed osteosarcoma cell migration and invasion. Moreover, introducing miR-193a-3p and miR-193a-5p mimics into MG63.2 cells or antagomiRs into MG63 cells confirmed their critical roles in osteosarcoma metastasis. Additionally, bioinformatics prediction along with biochemical assay results clearly suggested that the secretory small GTPase Rab27B and serine racemase (SRR) were direct targets of miR-193a-3p and miR-193a-5p, respectively. These two targets are indeed involved in the miR-193a-3p- and miR-193a-5p-induced suppression of osteosarcoma cell migration and invasion. MiR-193a-3p and miR-193a-5p play important roles in osteosarcoma metastasis through down-regulation of the Rab27B and SRR genes and therefore may serve as useful biomarkers for the diagnosis of osteosarcoma and as potential candidates for the treatment of metastatic osteosarcoma.

JMJD5 is a potential oncogene for colon carcinogenesis.

OBJECTIVE: To observe the effects of Jumonji C domain-containing (JMJD) 5 depletion on colon cancer (CC). METHODS: A short-hairpin RNA targeting JMJD5 was transfected into a lentivirus to make Lv-shJMJD5 for infection into the Caco-2 human cell. Besides, a negative control shRNA was constructed. The mRNA and protein levels of JMJD5 were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, respectively. Cell proliferation, migration, and invasion were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), soft agar colony assay and transwell assay, respectively. In addition, immunohistochemical (IHC) staining was performed to investigate the expression of JMJD5 in adjacent normal tissues and tumor tissues from patients with CC. RESULTS: Compared with control group, mRNA and protein levels of JMJD5 was significantly reduced after infection with Lv-shJMJD5 (P<0.05), and Caco-2 cell proliferation, migration, and invasion were all obviously inhibited (P<0.05). The results of IHC showed that JMJD5 was significantly up-regulated compared with normal tissues (P<0.01). Additionally, follow-up data demonstrated that the survival rate of patients with high expression of JMJD5 was obviously lower than that with low expression (P<0.01). CONCLUSIONS: JMJD5 depletion could significantly inhibit human CC cell proliferation, migration, and invasion, implying that JMJD5 might be a potential oncogene.

[Value of peritoneoscopy via natural orifice transluminal endoscopic surgery in the diagnosis of peritoneal carcinomatosis].

OBJECTIVE: To evaluate the value of peritoneoscopy via natural orifice transluminal endoscopic surgery (NOTES) in the diagnosis of patients with peritoneal carcinomatosis. METHODS: A total of 32 patients with peritoneal carcinomatosis were diagnosed by histological examination of biopsies at our hospital from April 2007 to October 2010. Their data of clinical manifestations, gastroscopy, colonoscopy, abdominal ultrasonography, abdominal computed tomography, magnetic resonance imaging, ascitic cytology and transgastric peritoneoscopy via NOTES were analyzed retrospectively. RESULTS: Among them, gastrointestinal cancers were diagnosed by digestive endoscopy in 9 cases (28.1%). And ovarian lesions in 8 cases (25.0%), pancreatic cancer in 2 cases (6.3%), primary liver cancer in 2 cases (6.3%) and bile duct carcinoma in 1 case (3.1%) were suspected according to imaging examinations. No peritoneal carcinomatosis was found by digestive endoscopy or imaging examinations. Ascitic cytology was positive in 6 cases (18.8%). Peritoneal carcinomatosis was diagnosed by transgastric peritoneoscopy via NOTES with histological examination of biopsies in all patients. Their findings of transgastric peritoneoscopy via NOTES were divided into 5 types, i.e., mass type (n = 3, 9.4%), nodular type (n = 5, 15.6%), ulcerative type (n = 1, 3.1%), omentum-embracing type (n = 1, 3.1%) and mixture type (n = 22, 68.8%). CONCLUSION: Transgastric peritoneoscopy via NOTES with histological examination of biopsies has important value in the pathologic diagnosis and the endoscopic typing of peritoneal carcinomatosis.