COVID-19 screening using breath-borne volatile organic compounds.

Rapid screening of COVID-19 is key to controlling the pandemic. However, current nucleic acid amplification involves lengthy procedures in addition to the discomfort of taking throat/nasal swabs. Here we describe potential breath-borne volatile organic compound (VOC) biomarkers together with machine learning that can be used for point-of-care screening of COVID-19. Using a commercial gas chromatograph-ion mobility spectrometer, higher levels of propanol were detected in the exhaled breath of COVID-19 patients (N= 74) and non-COVID-19 respiratory infections (RI) (N= 30) than those of non-COVID-19 controls (NC)/health care workers (HCW) (N= 87), and backgrounds (N= 87). In contrast, breath-borne acetone was found to be significantly lower for COVID-19 patients than other subjects. Twelve key endogenous VOC species using supervised machine learning models (support vector machines, gradient boosting machines (GBMs), and Random Forests) were shown to exhibit strong capabilities in discriminating COVID-19 from (HCW + NC) and RI with a precision ranging from 91% to 100%. GBM and Random Forests models can also discriminate RI patients from healthy subjects with a precision of 100%. In addition, the developed models using breath-borne VOCs could also detect a confirmed COVID-19 patient but with a false negative throat swab polymerase chain reaction test. It takes 10 min to allow an entire breath test to finish, including analysis of the 12 key VOC species. The developed technology provides a novel concept for non-invasive rapid point-of-care-test screening for COVID-19 in various scenarios.

Co-ablation versus cryoablation for the treatment of stage III-IV non-small cell lung cancer: A prospective, noninferiority, randomized, controlled trial (RCT).

BACKGROUND: This study compared a co-ablation (CA) system, which is a novel ablation device, with an argon-helium cryoablation (AHC) system. We aimed to compare the efficacy and safety of CA and AHC for the treatment of stage III-IV non-small cell lung cancer (NSCLC). METHODS: We conducted a multicenter randomized controlled trial (RCT) to determine whether CA was noninferior to AHC. The primary efficacy endpoints were the iceball coverage rate (ICR) and the disease control rate (DCR) one month after treatment. Noninferiority was declared if the lower limit of two-sided 95% confidence interval (CI) was less than 10%. The ICR and DCR were identified by logistic regression. Treatment safety was assessed. RESULTS: A total of 81 patients underwent randomization (41 assigned to the CA and 40 assigned to the AHC groups)and transthoracic ablation. The ICRs in the CA and AHC groups were 99.24% ± 2.18% and 98.66% ± 3.79%, respectively. Central lesions were associated with an increased risk of an incomplete ICR. The DCRs in the CA and AHC groups were 97.6% and 95%, respectively. A smaller lesion area in the CA group was significantly correlated with a better DCR. The rate of complications was 29.26% in the CA group and 30% in the AHC group. (P = 0.943). There was less probe usage per patient in the CA group. CONCLUSIONS: We determined that CA is noninferior to AHC in terms of efficacy and safety for the treatment of stage III-IV NSCLC. A smaller lesion area in the CA group was significantly correlated with a better DCR. KEY POINTS: CA was noninferior to AHC for stage III-IV NSCLC.

Hsa_circ_0046159 is involved in the development of chronic thromboembolic pulmonary hypertension.

The present study was performed to screen for potential molecular biomarkers and to assess the underlying mechanisms of chronic thromboembolic pulmonary hypertension (CTEPH) by using sequencing data analysis of microRNAs (miRNAs) and circular RNAs (circRNAs). Total RNA was isolated from peripheral-blood samples from five CTEPH patients and from five normal individuals. Based upon the identification of differentially expressed miRNAs (Affymetrix miRNA chip) and circRNAs (Agilent circRNA chip), target predictions for these differentially expressed miRNAs and functional enrichment analyses of the miRNAs and circRNAs were performed. Subsequently, the miRNA partner predictions of these differentially expressed circRNAs and co-expression analyses of differentially expressed circRNAs and miRNAs were conducted. Based on the results of these analyses, a competing endogenous RNA (ceRNA) network was constructed. Finally, the expression of circRNAs was detected by quantitative real-time PCR (qRT-PCR). Within the miRNA-circRNA regulatory network, hsa_circ_0026480 and hsa_circ_0046159 were predicted to interact with miR-27a-3p and miR-1226-3p, respectively with greater degree. Specially, ATP2A2-that had a ceRNA relationship with hsa_circ_0046159-was predicted as a target of miR-1226-3p. The results of RT-PCR also revealed a significantly increased expression of hsa_circ_0046159 in CTEPH samples than that in normal samples.

Mesenchymal stem cells overexpressing heme oxygenase-1 ameliorate lipopolysaccharide-induced acute lung injury in rats.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common and potentially lethal clinical syndromes characterized by acute respiratory failure resulting from excessive pulmonary inflammation, noncardiogenic pulmonary edema, and alveolar-capillary barrier disruption. At present, there is no effective and specific therapy for ALI/ARDS. Mesenchymal stem cells (MSCs) have well-known therapeutic potential in patients with ALI/ARDS. Heme oxygenase-1 (HO-1), a cytoprotective enzyme, possesses antioxidative, anti-inflammatory, and antiapoptotic effects. Thus, a combination of MSC transplantation with HO-1 delivery may have an additional protective effect against ALI/ARDS. This study investigated the effect of HO-1-modified bone-marrow-derived MSCs (MSCs-HO-1) on lipopolysaccharide (LPS)-induced ALI and its underlying mechanisms. We established MSCs-HO-1 through lentiviral transduction. The ALI rat model was established by successive LPS inhalations following injection with MSCs-HO-1. The survival rate, histological changes in the lungs, total protein concentration and neutrophil counts in bronchoalveolar lavage fluid, lung wet/dry weight ratio, cytokine levels in serum and lungs, nuclear transcription factor-κB activity, and protein expression of Toll-like receptor 4 signaling adaptors were examined. Furthermore, the cell viability, apoptosis, and paracrine activity of MSCs-HO-1 were examined under inflammatory stimuli in vitro. MSCs-HO-1 injection improved these parameters compared with primary unmodified MSCs. Moreover, MSCs-HO-1 had superior prosurvival and antiapoptotic properties and enhanced paracrine functions in vitro. Therefore, MSCs-HO-1 exert an enhanced protective effect to alleviate LPS-induced ALI in rats, and the mechanisms may be partially associated with superior prosurvival, antiapoptosis, and enhanced paracrine functions of MSCs-HO-1. These findings provide a novel insight into MSC-based therapeutic strategies for treating ALI/ARDS.

Overexpression of B7-H3 correlates with aggressive clinicopathological characteristics in non-small cell lung cancer.

Previous studies have investigated the prognostic significance of B7 homolog 3 (B7-H3) in non-small cell lung cancer (NSCLC), however, the results remain controversial. This study was aimed to determine the correlation between B7-H3 and survival as well as clnicalpathological characteristics in NSCLC using meta-analysis. We searched the electronic databases of PubMed, Embase, Web of Science, and China National Knowledge Infrastructure (CNKI) for relevant studies up to October 9, 2016. Pooled hazard ratios (HRs) and 95% confidence intervals (CIs) were used to estimate the impact of B7-H3 on overall survival (OS). Combined odds ratios (ORs) and 95%CIs were utilized to evaluate the correlations between B7-H3 and clinicalpathological features. This meta-analysis finally included 7 studies with 864 patients. The results showed that B7-H3 had no significant association with OS (HR=0.88, 95%CI: 0.36-2.13, p=0.776). High B7-H3 expression was a significant indicator of lymph node metastasis (OR=3.92, 95%CI: 2.65-5.81, p<0.001), and advanced TNM stage (OR=3.53, 95%CI: 2.45-5.09, p<0.001). B7-H3 has the potential to serve as a marker of tumor aggressiveness and lymph node metastasis in NSCLC. However, due to several limitations, further large-scale studies are needed to validate our results.

The Application of Cytidyl Guanosyl Oligodeoxynucleotide Can Affect the Antitumor Immune Response Induced by a Combined Protocol of Cryoablation and Dendritic Cells in Lewis Lung Cancer Model.

BACKGROUND Recently, several combined therapeutic strategies and targeted agents have been under investigation for their potential role in lung cancer. The combined administration of dendritic cells (DCs) and immune-adjuvant cytidyl guanosyl oligodeoxynucleotide (CpG-ODN) after cryosurgery has proven to be an effective strategy for treating lung cancer. However, whether the application of CpG-ODN could affect the therapeutic results remained to be further explored. MATERIAL AND METHODS The Lewis lung cancer (LLC)-bearing mice received cryoablation and injection of ex vivo-cultured DCs into the peritumoral zone. Subsequently, CpG-ODN was administered to experimental animals 6 hours, 12 hours, and 24 hours after DC injection. The mice in the control group received coadministration of DCs and CpG-ODN simultaneously. Therapeutic effects were evaluated by survival rates. The resistance to rechallenge of LLC cell was assessed by lung metastasis and in vitro cytotoxicity of splenocytes. Furthermore, T-cell subsets and multiple cytokines (interleukin [IL]-4, -10, and-12; interferon [IFN]-γ; tumor necrosis factor [TNF]-α) in the blood were assessed to elucidate the underlying mechanisms. RESULTS Higher ratios of CD4+ and CD8+ T cells and higher levels of IL-12, IFN-γ, and TNF-α were found in the blood of the mice that received CpG-ODN therapy 12 h after DC injection. The cytotoxicity potency of the splenocytes of these mice was significantly higher compared with the mice in other groups. Moreover, the mice receiving CpG-ODN therapy 12 h after DC injection showed significantly better resistance to rechallenge. Compared with the mice in other groups, the mice receiving CpG-ODN therapy 12 h after DC injection were superior in survival rates and antimetastatic effects. CONCLUSIONS Our study suggested that the therapeutic efficacy was closely associated with CpG-ODN administration in the combined therapeutic protocol of cryoablation, DCs, and immune adjuvant. In situ administration of CpG-ODN 12 h after DC injection might be considered the optimum application.

siRNA delivered by EGFR-specific scFv sensitizes EGFR-TKI-resistant human lung cancer cells.

The overexpression of epidermal growth factor receptor (EGFR) is closely associated with a poor outcome in non-small cell lung cancer (NSCLC), and EGFR is an ideal biomarker for the targeted therapy of NSCLC. Although patients with EGFR-activating mutations respond to EGFR tyrosine kinase inhibitors (EGFR-TKIs), they eventually acquire resistance, which typically results from a secondary EGFR mutation or the activation of other signaling pathways. Novel approaches to overcome or prevent EGFR-TKI resistance are clinically important. In this study, we developed an EGFR-scFv-arginine nonamer peptide fusion protein, s-9R, as an siRNA carrier. Here, we show that s-9R effectively and specifically delivers EGFR-siRNAs, KRAS-siRNA and MET-siRNA into NSCLC cells and silences the expression of target genes. The sensitivity of NSCLC cells to gefitinib was restored after treatment with the s-9R/siRNA complex, and the apoptosis rates of the treated cells were significantly higher than those of the control groups. Furthermore, the co-administration of s-9R/siRNA and gefitinib successfully suppressed the progression of H1975 xenograft tumors and extended the life span of tumor-bearing nude mice. Collectively, the results of this study provide not only a new scFv derivative for delivering siRNA into EGFR-overexpressing, TKI-resistant NSCLC cells but also a novel method for overcoming TKI resistance.

Multicentre investigation of pathogenic bacteria and antibiotic resistance genes in Chinese patients with acute exacerbation of chronic obstructive pulmonary disease.

OBJECTIVE: A prospective observational study to investigate the distribution and antimicrobial resistance of pathogenic bacteria in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in Beijing, China. METHODS: Patients with AECOPD were recruited from 11 general hospitals. Sputum specimens were cultured and bacteria identified. Antibiotic susceptibility was determined for each isolate, and presence of antibiotic resistance genes was evaluated using polymerase chain reaction. RESULTS: Pathogenic bacteria were isolated from 109/318 patients (34.28%); 124 isolates of 22 pathogenic bacterial species were identified, including Klebsiella pneumoniae (16.94%), Pseudomonas aeruginosa (16.94%), Acinetobacter baumannii (11.29%), Streptococcus pneumoniae (8.87%), and Staphylococcus aureus (7.26%). S. aureus was sensitive to tigecycline, teicoplanin, vancomycin and linezolid but resistant to penicillin and levofloxacin. K.pneumoniae, P. aeruginosa, A. baumannii and E. coli were susceptible to amikacin and cefoperazone. CONCLUSIONS: K. pneumoniae and P. aeruginosa are the most common pathogenic bacteria in AECOPD cases in Beijing, China. Our antibiotic resistance findings may be helpful in selecting antibiotic therapy.

Therapeutic Potential of Umbilical Cord Mesenchymal Stem Cells for Inhibiting Myofibroblastic Differentiation of Irradiated Human Lung Fibroblasts.

Radiation-induced lung injury (RILI) limits the benefits of radiotherapy in patients with lung cancer. Radiation-induced differentiation of lung fibroblasts to myofibroblasts plays a key role in RILI. Recent studies have shown that mesenchymal stem cells (MSCs) can protect against lung fibrosis and that Wnt/ß-catenin signaling is involved in fibrotic processes. In the present study, we explored the therapeutic potential of human umbilical cord MSCs (HUMSCs) for preventing radiation-induced differentiation of human lung fibroblasts (HLFs) to myofibroblasts. There are two advantages in the use of HUMSCs; namely, they are easily obtained and have low immunogenicity. Irradiated HLFs were co-cultured with HUMSCs. Expression of α-smooth muscle actin (α-SMA), a myofibroblast marker, was measured by Western blot analysis and immunohistochemistry. Irradiation (X-rays, 5 Gy) induced the differentiation of HLFs into myofibroblasts, which was inhibited by co-culture with HUMSCs. Irradiation also caused activation of the canonical Wnt/ß-catenin signaling in HLFs, as judged by increased phosphorylation of glycogen synthase kinase 3ß, nuclear accumulation of ß-catenin, and elevated levels of Wnt-inducible signaling protein-1 (WISP-1) in the conditioned medium. However, co-culture with HUMSCs attenuated the radiation-induced activation of the Wnt/ß-catenin signaling. We also measured the expression of FRAT1 that can enhance the Wnt/ß-catenin signaling by stabilizing ß-catenin. Co-culture with HUMSCs decreased FRAT1 protein levels in irradiated nHLFs. Thus, co-culture with HUMSCs attenuated the radiation-induced activation of Wnt/ß-catenin signaling in HLFs, thereby inhibiting myofibroblastic differentiation of HLFs. Wnt/ß-catenin signaling is a potential therapeutic target for limiting RILI in patients receiving radiotherapy for lung cancer.

[Effect of irradiated human lung fibroblasts on activation of canonical Wnt/ß-catenin signaling pathway in mesenchymal stem cells].

OBJECTIVE: To investigate the effect of irradiated human lung fibroblasts (HLFs) on the canonical Wnt/ß-catenin signaling pathway in human umbilical cord mesenchymal stem cells (HUMSCs). METHODS: HUMSCs were cultured alone (single group) or co-cultured with HLFs exposed to 5Gy X-rays (co-culture group). The protein levels of GSK-3ß, p-GSK-3ß, FRAT1 and ß-catenin in HUMSCs were examined by Western blotting 3 days after culture or co-culture. WISP-1 protein levels in conditioned medium were examined by ELISA. RESULTS: The levels of p-GSK3ß/GSK3ß (0.15 ± 0.05), FRAT1 (0.48 ± 0.07) and ß-catenin (0.50 ± 0.07) in co-cultured HUMSCs significantly decreased compared to those in single group (0.55 ± 0.05, 1.16 ± 0.13 and 2.39 ± 0.15, all P<0.05). The supernatant level of WISP-1 in co-culture group was significantly decreased [(602.23 ± 161.47) ng/mL], compared to the single group [(977.77 ± 110.56) ng/mL, P<0.05]. CONCLUSION: Irradiated HLFs attenuate the activation of canonical Wnt/ß-catenin signaling pathway in HUMSCs in vitro.

[Evaluation of bronchial mucosa involvement in sarcoidosis patients using ¹8F-FDG PET-CT].

OBJECTIVE: To explore the value of ¹8F-FDG PET-CT in evaluating bronchial mucosa involvement in patients with saroidosis. METHODS: A retrospective analysis was conducted among 6 sarcoidosis patients with and 14 patients without bronchial mucosa involvement to collect the data including the standard uptake value (SUVMax/Mean) of ¹8F-FDG, serum angiotensin converting enzyme (sACE), and proportion of lymphocytes and CD4⁺/CD8 ⁺ T lymphocyte ratio in bronchoalveolar lavage fluid (BALF). RESULTS: The lung focal SUV(Max/Mean) was higher in patients with bronchial mucosa involvement than those without (7.04 ± 5.83/5.00 ± 4.69 vs 5.68 ± 3.66/3.82 ± 2.39), but such differences were not statistically significant (P=0.565/0.495). The SUV(Max/Mean) of the hilum of the lung and the mediastina lymph nodes were significantly higher in patients with bronchial mucosa involvement (13.28 ± 5.57/10.48 ± 4.43 vs 6.20 ± 1.77/4.52 ± 1.43, P=0.0003/0.0002; 13.84 ± 4.35/9.69 ± 2.74 vs 7.16 ± 2.52/5.28 ± 1.77, P=0.0004/0.0004). The level of sACE and CD4⁺/CD8 ⁺ T lymphocyte ratio in BALF were also significantly higher in patients with bronchial mucosa involvement (60.58 ± 16.3 vs 49.16 ± 13.3 IU/L, P=0.045; 7.30 ± 5.0 vs 2.90 ± 3.1, P=0.026). The proportion of lymphocytes in BALF was comparable between the patients with and without bronchial mucosa involvement (44.10 ± 10.3% vs 35.30 ± 12.5%, P=0.148). CONCLUSIONS: For patients with saroidosis, ¹8F-FDG PET-CT is useful in evaluating bronchial mucosa involvement, which is one of the key features of active sarcoidosis.

AC-NP: a novel chimeric peptide with natriuretic and vasorelaxing actions.

The aim of this study was to evaluate the cardiovascular and renal activities of a newly designed natriuretic peptide (NP). Here, we engineered a novel 28-amino acid chimeric peptide, termed AC-NP that combined the 17-amino acid ring of C type natriuretic peptide (CNP) with the 6-amino acid N-terminus and 5-amino acid C-terminus of atrial natriuretic peptide (ANP). Both in vitro and in vivo experiments were performed to determine the actions of AC-NP. In normal rats, AC-NP proved to be more potentially diuretic, natriuretic and hypotensive compared with other NPs, such as ANP, CNP and vasonatrin peptide (VNP), which is another man-made NP. In relaxation of isolated abdominal aorta from rat, AC-NP was equally effective to ANP, CNP and VNP. Elevated levels of 3',5'-guanosine monophosphate (cGMP) in plasma and urine cGMP excretion indicated the participation of cGMP in the functions of AC-NP. Taken together, innovative designed AD-NP might be a new candidate therapeutic peptide against cardiorenal disorders.

Protective effects of vasonatrin peptide against hypobaric hypoxia-induced pulmonary hypertension in rats.

1. The aim of the present study was to investigate the in vivo effects of vasonatrin peptide (VNP) on hypoxia-induced pulmonary hypertension (HPH). 2. The HPH model was developed by subjecting rats to hypobaric hypoxia. The HPH rats were then treated with either VNP (50 microg/kg per day, i.p.) or saline (0.5 mL, i.p.) every day for 7 days. Haemodynamic indices, right ventricular hypertrophy (RVH) and remodelling of the pulmonary arteries were evaluated. In addition, plasma levels of atrial natriuretic peptide (ANP), endothelin (ET)-1 and angiotensin II (AngII) were determined, as was natriuretic peptide receptor-C (NPR-C) mRNA expression in the right ventricle. 3. Hypobaric hypoxia induced severe HPH compared with the normoxic control group. Treatment of HPH rats with VNP for 1 week significantly reduced mean pulmonary arterial pressure, pulmonary vascular resistance, RVH and muscularization of the pulmonary arteries, although pulmonary blood flow was increased in this group. In addition, significantly lower levels of plasma ET-1 and AngII and cardiac NPR-C mRNA expression were observed in VNP-treated compared with saline-treated HPH rats, whereas higher plasma concentrations of ANP were found in the former group. Acute intravenous administration of 50 microg/kg VNP significantly ameliorated pulmonary haemodynamics in HPH rats. 4. Taken together, the date indicate that VNP has certain preventative and therapeutic effects against HPH.

[Maintenance chemotherapy in 43 patients with advanced non-small cell lung cancer: a retrospective study.].

BACKGROUND: It is unclear that patients with advanced non-small cell lung cancer (NSCLC) should receive prolonged chemotherapy or the best support, after achieving a maximum tumor response with 4 to 6 initial chemotherapy cycles. The aim of this study is to compare maintenance paclitaxel therapy with observation in advanced NSCLC patients. METHODS: A total of 166 patients with stage IIIB or IV NSCLC were treated with four monthly paclitaxel-carboplatin as following: paclitaxel 100 mg/m(2) weekly for 3 of 4 weeks with carboplatin (area under the curve[AUC]=6) on day 1. Patients who responded at week 16 were assigned to either weekly paclitaxel therapy(70 mg/m(2), 3 of 4 weeks) or observation. Maintenance therapy continued until disease progression, development of intercurrent illness, intolerable toxicity, or investigator decision to terminate treatment. RESULTS: There was a longer median time to progression in maintenance therapy compared with observation (P< 0.05). One- and two-year survival rates were 65% and 24% in the paclitaxel arm and 59% and 20% in the observation arm, respectively (P> 0.05). The performance status and the stage were the independent prognostic factors by Cox regression. CONCLUSIONS: Maintenance paclitaxel could benefit median time to progression but not survival time in patients who responded to induction chemotherapy. Nevertheless, further evaluation of maintenance therapy should be done before the concept is abandoned.

Thoracic masses treated with percutaneous cryotherapy: initial experience with more than 200 procedures.

PURPOSE: To perform and report initial experience with percutaneous cryotherapy (PCT) of the thorax. MATERIALS AND METHODS: A human investigation committee approved the study protocol, and all patients gave informed consent. One hundred eighty-seven patients who were not surgical candidates underwent computed tomography (CT)-guided PCT for treatment of thoracic cancer masses. CT-visualized low-attenuating ice formation after PCT was compared with initial tumor size and location. At 1 week and at 1, 3, 6, and 12 months after PCT, the various findings seen on available CT scans and any complications were noted. chi(2) and Student t tests were used to identify significant differences in frequencies and mean values of imaging observations, respectively. RESULTS: Ice formation was identified at CT as reduced attenuation values (in Hounsfield units) within soft-tissue masses, the mean sizes of which were 4.3 cm +/- 0.2 (standard deviation) in peripheral locations and 6.4 cm +/- 0.3 in central locations. Tumor size and location were independent predictors of tumor coverage by low-attenuating ice: Mean coverage was 99% for peripheral masses 4 cm or smaller (n = 101) and 80% for central masses larger than 4 cm (n = 58) (P < .001). An area of necrotic cavitation larger than the original mass developed in 80% (77 of 96) of masses within 1 week and was nearly resolved by 3 months in 7% (five of 76) of masses. By 6 months, minimal pulmonary scarring was noted in 56 patients and 86% of masses showed reduced or stable size. The overall rate of pneumothorax was only 12% (22 of 187 patients), and other side effects appeared to be self limited. No major bleeding or bronchial damage was noted. Two deaths in debilitated patients were temporally related, and two complications involved brachial and recurrent laryngeal nerve damage. The patient with laryngeal nerve damage regained speech within 2 months. CONCLUSION: CT-guided PCT yielded low procedural morbidity given the extent of freezing, even near mediastinal structures. Ongoing advances in cryotechnology, imaging guidance, and treatment planning may help to avoid the degree of undertreatment of larger central masses observed in this study.

Targeting ischemic stroke with a novel opener of ATP-sensitive potassium channels in the brain.

During cerebral ischemia, the opening of neuronal ATP-sensitive potassium channels (K(ATP) channels) affords intrinsic protection by regulating membrane potential. To augment this endogenous mechanism, we have synthesized iptakalim, a K(ATP) opener. Through K(ATP) channel activation, iptakalim affected multiple pathways of the glutamatergic system, limiting glutamate release and receptor actions, which are involved in excitotoxicity during ischemic damage. The molecule readily penetrated the brain and showed low toxicity in animal experiments. In different animal models of stroke as well as in cell cultures, iptakalim provided significant neuroprotection, not only in promoting behavioral recovery but also in protecting neurons against necrosis and apoptosis. This compound thus has promise as a neuroprotective drug for the treatment of stroke and other forms of neuronal damage.