Values of different specimen preparation methods for the diagnosis of lung cancer by endobronchial ultrasound guided transbronchial needle aspiration.

BACKGROUND: Endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) has been become an important procedure for the diagnosis and staging of lung cancer. Our research identified the effects of different pathological preparation on the diagnosis of lung cancer for specimens obtained by biopsy. METHODS: Patients were clinically considered if lung cancer was accompanied by mediastinal or hilar lymph node enlargement between March 2014 and November 2017. Specimens obtained by EBUS-TBNA were treated by three methods: traditional smear cytology, liquid-based cytology (LBC) and histopathology. RESULTS: Of a total of 154 puncture sites from 153 patients, the total positive rate of combination for the three pathological treatment types (histopathology, direct traditional smear, and LBC) was 77.3%. The diagnostic positive rate for histopathology was 68.6%, direct traditional smear was 65.6%, and LBC was 60.4%; there was no significant differences among the three single pathological treatment types (P = 0.29), but there was a statistically significant difference between the combination of three treatments and any single pathological treatment type (P = 0.01). The diagnostic sensitivities of histopathology combined with traditional smear and histopathology combined LBC were 94.4 and 92.8%, respectively, the specificities and PPVs were both 100%, and the diagnostic accuracies were 95.5 and 94.2%, respectively; the sensitivities, specificities and diagnostic accuracies above were all higher than those of single specimen treatment and lower than those of the three combined. CONCLUSION: When EBUS-TBNA is used for the diagnosis and staging of lung cancer, the use of histopathological sections combined with direct cytological smear should be sufficient and is the most economical choice.

A positive feedback loop between ZNF205-AS1 and EGR4 promotes non-small cell lung cancer growth.

Accumulating evidences revealed that long noncoding RNAs (lncRNAs) are frequently implicated in non-small cell lung cancer (NSCLC). Herein, we reported the identification of a novel NSCLC-associated functional lncRNA ZNF205 antisense RNA 1 (ZNF205-AS1). ZNF205-AS1 was increased in NSCLC tissues and cell lines, and associated with poor prognosis of NSCLC patients. Bioinformatics prediction, combined with experimental verification revealed that early growth response 4 (EGR4) directly bound to ZNF205-AS1 promoter, increased the promoter activity of ZNF205-AS1, and activated ZNF205-AS1 transcription. Intriguingly, ZNF205-AS1 transcript directly interacted with EGR4 mRNA, increased EGR4 mRNA stability, and up-regulated EGR4 expression via RNA-RNA interaction. Thus, ZNF205-AS1 and EGR4 formed a positive feedback loop. Through regulating EGR4, ZNF205-AS1 activated its own promoter activity. EGR4 was also increased in NSCLC and the expression of ZNF205-AS1 was significantly positively correlated with EGR4 in NSCLC tissues. Gain-of-function and loss-of-function assays demonstrated that both ZNF205-AS1 and EGR4 promoted NSCLC cell growth in vitro and NSCLC tumour growth in vivo. Concurrently depleting ZNF205-AS1 and EGR4 more significantly repressed NSCLC tumour growth in vivo. Collectively, our study demonstrated that the positive feedback loop between ZNF205-AS1 and EGR4 promotes NSCLC growth, and implied that targeting this feedback loop may be promising therapeutic strategy for NSCLC.

The miRNA hsa-miR-6515-3p potentially contributes to lncRNA H19-mediated-lung cancer metastasis.

Aberrant expression of long noncoding RNAs (lncRNAs) contributes to all phenotypes of cancer including metastasis, which is a major cause of death in many advanced malignancies. One particular lncRNA, H19, is found to be a crucial player in cancer progression by modulating multiple microRNAs (miRNAs). In this study, we screened miRNAs possibly associated with H19 using lung carcinoma cell lines and patient with lung cancer tissues, and selected one possible hit, hsa-miR-6515-3p, to perform in vitro functional assays. Its inhibition leads to decreased proliferation and migration of SPC-A1 lung cancer cells and is in good correlation with H19-knockdown groups. These results indicate that H19 may be an epigenetic regulator of miR-6515-3p, and its dysregulation may contribute to lung cancer progression and metastasis.

A rare case of an HIV-seronegative AIDS patient with Pneumocystis jirovecii pneumonia.

BACKGROUND: As technology progresses, several highly sensitive human immunodeficiency virus (HIV) screening kits are being researched and developed to quickly and efficiently identify serum HIV antibodies within the non-window period. In individuals who are HIV-seronegative, HIV infections that are not within a window period are rare. In such cases, all antibody detection methods will fail, and misdiagnosing these patients will have catastrophic consequences. CASE PRESENTATION: A 22-year-old male Chinese patient with diffuse exudative lesions in both lungs and initial symptoms of cough and dyspnoea was diagnosed with Pneumocystis jirovecii pneumonia (PJP) by aetiological examination, and the patient's plasma CD4+ T-cell count was extremely low. In China, PJP is prevalent in HIV-infected individuals. Pneumocystis jirovecii (P. jirovecii) has a high colonisation rate in patients with HIV infections. This patient was naturally suspected of being an HIV patient; however, serum HIV antibody tests were negative using both an enzyme-linked immunosorbent assay (ELISA) and a latex agglutination assay, and HIV was not detected by western blotting. Subsequently, the plasma HIV viral load was found to be extremely high on two repeated plasma HIV RNA tests, thus confirming HIV-seronegative acquired immunodeficiency syndrome (AIDS) in this patient. With administration of effective anti-P. jirovecii treatment and highly active antiretroviral therapy (HAART) after diagnosis, the patient's disease condition was rapidly controlled. CONCLUSION: This is the second reported case in China of an HIV-seronegative AIDS patient. Such cases are also rare worldwide. Although HIV-seronegative HIV infections are rare, AIDS should be considered in immunodeficient patients with opportunistic infections, even if the test results are HIV-seronegative. Plasma HIV RNA testing is important for such patients.

Suppression of ZNF205-AS1/EGR4 positive feedback loop attenuates cisplatin resistance of non-small cell lung cancer cells via targeting miR-138-5p/OCT4 pathway.

Background: Long non-coding RNAs (lncRNAs) are frequently reported to involve in the onset and development of non-small cell lung cancer (NSCLC). Cisplatin (DDP) resistance continues to pose a daunting challenge for improving the prognosis of NSCLC patients. The current study intends to elucidate the molecular mechanisms underlying the function of lncRNA ZNF205 AS1/early growth response 4 (EGR4) positive feedback loop in DDP resistance of NSCLC. Methods: A series of assays, including real-time polymerase chain reaction (PCR), western blotting, flow cytometry, and dual-luciferase reporter, were performed to evaluate the effect of ZNF205-AS1/EGR4 loop in the established DDP-resistant A549 cell line and its progenitor A549 cell line. Immunohistochemistry (IHC) technique was conducted to investigate the expression pattern of EGR4 and octamer-binding protein 4 (OCT4) in NSCLC tissues. RNA pull-down assay was carried out to evaluate the interaction between miR-138-5p and EGR4 and OCT4. Transwell assay and wound healing assay was used to evaluate the invasive and migratory potential of cells subject to various treatment. The protein levels of Bcl2, Bax, Cl-caspase 3, Cl-PARP and OCT4 were measured in western blotting assay. Results: The levels of ZNF205-AS1, EGR4 and OCT4 were notably upregulated in post-chemotherapy DDP-resistant lung specimens, as opposed to those pre-chemotherapy, and in A549/DDP cells than the progenitor DDP-sensitive A549 cells. In contrast, the level of miR-138-5p was significantly reduced in A549/DDP cells (P<0.05). Luciferase reporter assay confirmed the interaction between ZNF205-AS1 and miRNA-138-5p. Protein-RNA interaction was validated between miR-138-5p, EGR4 and OCT4. The higher chemosensitivity of DDP-resistant cells induced by the loss-of-function of ZNF205-AS1 could be diminished by a miR-138-5p inhibitor. Conclusions: Our data demonstrated that miR-138-5p/OCT4 functions as a downstream effector of the ZNF205-AS1/EGR4 positive feedback loop and mediates resistance of NSCLC cells to DDP. Our work sheds light on the therapeutic strategies for NSCLC with DDP chemoresistance.

Sintilimab plus anlotinib as second- or third-line therapy in metastatic non-small cell lung cancer with uncommon epidermal growth factor receptor mutations: A prospective, single-arm, phase II trial.

BACKGROUND: Patients with non-small-cell lung cancer (NSCLC) and uncommon EGFR alterations typically have worse treatment outcomes than patients with classically EGFR-mutated NSCLC. This study aimed to investigate the efficacy and safety of PD-1 blockade with sintilimab plus anti-angiogenic treatment with anlotinib in patients with NSCLC harboring uncommon EGFR mutations. METHODS: Patients with metastatic NSCLC harboring uncommon EGFR mutations after two previous treatments, including a platinum-based chemotherapy regimen and a targeted treatment (or chemotherapy only for patients harboring EGFR ex20ins), received sintilimab combined with anlotinib. The primary endpoint was objective response rate (ORR). RESULTS: At data cutoff (September 27, 2022), median follow-up was 22.3 months (range, 1.2-37.6). Among 21 enrolled patients, 12 had EGFR ex20ins and nine had other uncommon EGFR mutations such as L861Q, G719A, and G709X. Overall, eight patients (38.1%) achieved an objective response, and 18 (85.7%) achieved disease control. Median (95% CI) progression-free survival (PFS) was 7.0 (5.4-8.6) months, and median overall survival (OS) was 20.0 (15.6-24.4) months. The 12-month PFS rate (95% CI) was 22.2% (7.4-42.0), and the 12-month OS rate was 66.7% (42.5-82.5). Patients harboring EGFR ex20ins had similar ORR and PFS to those with other mutations. Six patients (28.6%) experienced grade 3 treatment-related adverse events (TRAEs); hand-foot syndrome was the most common grade 3 TRAE (2 patients; 9.5%). No grade ≥4 TRAEs were observed. CONCLUSIONS: The combination of sintilimab and anlotinib demonstrated durable efficacy and was generally well tolerated in patients with NSCLC and uncommon EGFR mutations who had received prior standard-of-care treatments. (ClinicalTrials.gov identifier: NCT04790409).

Hypoxia upregulates the expression of lncRNA H19 in non-small cell lung cancer cells and induces drug resistance.

Background: The incidence of lung cancer is extremely high. For treatment, the chemotherapy of lung cancer is low, and drug resistance and recurrence are frequently observed, in which hypoxic tumor microenvironments play a key role. We investigated the upregulation of long non-coding ribonucleic acid (lncRNA) H19 expression and the induction of drug resistance in non-small cell lung cancer (NSCLC) cells under hypoxic conditions. Methods: We used cobaltous chloride (CoCl2) to create hypoxic culture environments for the A549 and H1650 cells, and they were divided into normoxia and hypoxia groups. We used real-time quantitative PCR detecting system (qPCR) to detect the messenger RNA (mRNA) expressions of lncRNA H19, hypoxia-inducible factor 1-α (HIF1-α), multi-drug resistant associated protein 1 (MRP1), and heme oxygenase-1 (HO-1), and western blot to detect the protein expressions of HIF1-α, MRP1, HO-1, and P-glycoprotein (P-gp). We performed cell migration and invasion assays. Annexin V-Allophycocyanin (APC) and flow cytometry were used to detect the apoptotic rates. Results: We found that the expression levels of lncRNA H19, HIF1-α, HO-1, multi-drug resistant associated protein 1 (MRP1), and P-glycoprotein increased significantly in the NSCLC cell lines (A549 and H1650) under hypoxic conditions, as determined by the qPCR and western blot analyses. In NSCLC cells cultured under hypoxic conditions, the invasion and migration ability of tumor cells increased significantly, resistance to cisplatin increased, and cisplatin-induced apoptosis decreased considerably. In addition, chemoresistance was also induced in tumor cells under hypoxic conditions. Conclusions: These results indicate that hypoxia increased the expression levels of lncRNA H19 and induced chemoresistance in tumor cells.

The removal of floating right heart thrombi and pulmonary embolus using AngioJet device and venoarterial extracorporeal membrane oxygenation: a case report.

Background: Floating right heart thrombi (FRHTS), known as thrombi in transit, are usually located in the atrium or ventricle. Generally, it occurs in patients with pulmonary embolism (PE) and dyspnea, chest pain, syncope and palpitations are the most common symptoms on presentation. The mortality of patients with FRHTS is higher than that of those without FRHTS. Current treatment includes anticoagulation, systemic thrombolysis, catheter directed interventions, and surgical embolectomy. However, there is no consensus on the optimal management options. Case Description: Herein, we report the case of a patient who presented with hypotension and tachycardia accompanied by an asymptomatic right leg deep vein thrombosis, right atrial thrombus, and pulmonary embolus. He had a history of radical resection of colon cancer 1 month prior. And he had developed chest tightness accompanied by stabbing pain in the chest area 1 day ago. He experienced an episode of syncope 8.5 hours ago. So he was referred to the local hospital. After the pulmonary computed tomography angiography (CTA) scan, he was diagnosed with pulmonary embolus and administrated with 5,000 u low molecular weight heparin. Then he was transferred to our hospital. On arrival in the emergency department, the bedside transthoracic echocardiography (TTE) revealed there was an enlarged right atrium and right ventricle, with a floating right atrial mass prolapsing through the tricuspid valve during diastole. The patient accepted anticoagulation treatment, but refused to undergo thrombolysis or surgical embolectomy. Eventually, the right heart thrombi (RiHT) floated to the left main branch of pulmonary artery. It was successfully treated by using AngioJet device and venoarterial extracorporeal membrane oxygenation (VA-ECMO). Our case provides clinical evidence supporting the feasibility and efficacy of AngioJet device and VA-ECMO in the treatment of the RiHT and PE. Conclusions: Patients with PE combined with RiHT have higher mortality than those without RiHT, VA-ECMO could be used to maintain the circulation, and the AngioJet device could be used as an alternative treatment for patients who are reluctant to receive thrombolysis or surgical embolectomy.

Concurrent infection with Talaromyces marneffei and Cryptococcus neoformans in a patient without HIV infection.

A case report of coinfection with Talaromyces marneffei (T. marneffei) and Cryptococcus neoformans (C. neoformans) is presented in a 57-year-old woman with hemolytic anemia who received dexamethasone for 8 years. To the best of our knowledge, this patient was successfully treated with voriconazole. This is the first case of T. marneffei and C. neoformans coinfection in a HIV-negative host. Clinicians should be aware of concomitant infection with T. marneffei and other pathogens in immunocompromised hosts. The current case report highlights the importance of clinician awareness of concurrent infections with T. marneffei and other pathogens in immunosuppressed patients.

Clinical analysis of 132 cases COVID-19 from Wuhan.

Numerous cases of pneumonia from a novel coronavirus (SARS-CoV-2) emerged in Wuhan, China during December 2019.We determined the correlations of patient parameters with disease severity in patients with COVID-19.A total of 132 patients from Wuhan Fourth Hospital who had COVID-19 from February 1 to February 29 in 2020 were retrospectively analyzed.Ninety patients had mild disease, 32 had severe disease, and 10 had critical disease. The severe/critical group was older (P < .05), had a higher proportion of males (P < .05), and had a greater mortality rate (0% vs 61.9%, P < .05). The main symptoms were fever (n = 112, 84.8%) and cough (n = 96, 72.7%). Patients were treated with antiviral agents (n = 94, 71.2%), antibiotics (n = 92, 69.7%), glucocorticoids (n = 46, 34.8%), intravenous immunoglobulin (n = 38, 27.3%), and/or traditional Chinese medicine (n = 40, 30.3%). Patients in the severe/critical group received mechanical ventilation (n = 22, 16.7%) or high-flow nasal can-nula oxygen therapy (n = 6, 4.5%). Chest computed tomography (CT) indicated bilateral pneumonia in all patients. Relative to the mild group, the severe/critical group had higher levels of leukocytes, C-reactive protein (CRP), procalcitonin (PCT), D-dimer, B-type natriuretic peptide (BNP), liver enzymes, and myocardial enzymes (P < .05), and decreased levels of lymphocytes and blood oxygen partial pressure (P < .05).The main clinical symptoms of patients from Wuhan who had COVID-19 were fever and cough. Patients with severe/critical disease were more likely to be male and elderly. Disease severity correlated with increased leukocytes, CRP, PCT, BNP, D-dimer, liver enzymes, and myocardial enzymes, and with decreased lymphocytes and blood oxygen partial pressure.

Bacterial etiology in early re-admission patients with acute exacerbation of chronic obstructive pulmonary disease.

BACKGROUND: Repeatedly hospitalized patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) are often exposed to more antibiotics, but the distribution of pathogenic bacteria in these patients is poorly understood. The objectives of this study were to analyze the distribution of pathogenic bacteria and the risk factors associated with multidrug-resistant (MDR) bacteria infection in early re-admission patients with AECOPD. METHODS: We retrospectively reviewed charts for patients with AECOPD admitted to our hospital between January 2011 and November 2012. The early re-admission group and non-early readmission group were determined by whether patients were readmitted within 31 days after discharge. Detection of potentially pathogenic microorganisms (PPMs) and MDR bacteria were analyzed. Logistic regression analysis was performed to identify independent risk factors for MDR bacteria infection. RESULTS: PPMs were isolated from 230 (32.0%) cases of respiratory tract specimens; MDR bacteria accounted for 24.7% (57/230). Pseudomonas aeruginosa (43.7%), Klebsiella pneumoniae (15.6%), and Acinetobacter baumannii (12.5%) were the top three PPMs in the early readmission group, while the top three PPMs in the non-early readmission group were K. pneumoniae (23.7%), P. aeruginosa (21.2%), and Streptococcus pneumoniae (17.1%). Multivariate analysis showed that use of antibiotics within 2 weeks (odds ratio [OR] 8.259, 95% confidence interval [CI] 3.056-22.322, p = 0.000) was the independent risk factor for MDR bacteria infection. CONCLUSION: Non-fermentative Gram-negative bacilli (NFGNB) and enterobacteria were the predominant bacteria in early readmission patients with AECOPD. The detection rate of MDR bacteria was high which was related to the use of antibiotics within 2 weeks before admission in these patients.

Shikonin­mediated inhibition of nestin affects hypoxia­induced proliferation of pulmonary artery smooth muscle cells.

The imbalance between the proliferation and apoptosis of pulmonary artery smooth muscle cells (PASMCs) is of importance in pulmonary vascular remodeling. Shikonin, a naphthoquinone compound extracted from the Chinese medicinal herb Lithospermum erythrorhizon, inhibits the proliferation of rat smooth muscle cells (SMCs). The present study was designed to investigate the effects of shikonin on the proliferation of rat PASMCs and the possible mechanisms involved. Rat PASMCs were cultured under the following five treatment conditions: Normal control; hypoxia for 24 h; hypoxia + 1 µM shikonin for 24 h; hypoxia + 2 µM shikonin for 24 h; and hypoxia + 4 µM shikonin for 24 h. The viability of PASMCs was measured using the Cell Counting Kit­8 assay, the mRNA expression of nestin (NES) in each group was measured by reverse transcription­polymerase chain reaction and the protein expression of NES was measured by western blotting. The proliferation of hypoxic PASMCs transfected with NES­specific small interfering (si)RNA decreased compared with the non­transfected group. These results indicated that hypoxia induced the proliferation of PASMCs through the enhancement of NES expression. The treatment of hypoxic PASMCs with shikonin resulted in a significant downregulation of NES expression and the inhibition of PASMC proliferation.

A rare fatal case of a novel bunyavirus-associated hemophagocytic lymphohistiocytosis.

Herein we describe a rare fatal case of a novel bunyavirus-associated hemophagocytic lymphohistiocytosis (HLH) in a 62-year-old female patient. The novel bunyavirus infects patients with or without HLH who have similar clinical features such as fever, thrombocytopenia, and leukocytopenia. Therefore, the diagnosis of HLH can be easily missed. When HLH occurs, the disease worsens and the fatality rate rises. Our finding highlights the importance of bone marrow biopsy performed as soon as possible for patients suspected of having a novel bunyavirus infection and showing marked cytopenia in three cell lines.

Pulmonary oncocytoma: a case report.

A 70-year-old man complained of no discomfort was admitted to our hospital because of a nodules was found in his lung. Chest computer tomography showed a nodules arising from the upper lobe of the left lung, and the lesion became larger in 2 years follow-up period. The patient underwent video-assisted thoracoscopic surgery of lobectomy to remove the nodules. Histologically, the tumor specimen contained multiple glandular structures with oncocytic cells lining. Immunohistochemical staining showed the tissue did not include some neuroendocrine granules. Finally, He was diagnosed as pulmonary oncocytoma. The patient discharged from our hospital after surgery and with no recurrence in 29 months period.

Risk factors associated with chronic obstructive pulmonary disease early readmission.

BACKGROUND: The 31 day readmission rate is deemed to be an important indicator of the quality of medical care in China. The objectives of this study were to identify the readmission rate of acute exacerbation for chronic obstructive pulmonary disease (COPD) and to evaluate associated risk factors. METHODS: We retrospectively reviewed charts for patients with acute exacerbation of COPD (AECOPD) admitted to our hospital between January 2011 and November 2012. The early-readmission group and non-early-readmission group were determined by whether patients were readmitted within 31 days after discharge. Logistic regression analysis was performed to identify risk factors for early readmission following an AECOPD. RESULTS: There were 692 patients with 925 admissions during the 23 month period; 63 (6.8%) admissions met our criteria for early readmission. Multivariate analysis showed that chronic cor pulmonale (odds ratio [OR] 2.14, 95% confidence interval [CI] 1.26-3.64, p = 0.005), hypoproteinemia (OR 2.02, 95% CI 1.03-3.95, p = 0.040) and an elevated PaCO2 (OR 1.03, 95% CI 1.00-1.06, p = 0.027) were identified as risk factors for early readmission of AECOPD. CONCLUSION: The readmission rate for AECOPD was 6.8%. AECOPD patients with chronic cor pulmonale, hypoproteinemia, and a high PaCO2 are at higher risk for readmission with 31 days of hospital discharge, and medical care of these patients warrants greater attention.