Progestogen-driven B7-H4 contributes to onco-fetal immune tolerance.

Immune tolerance mechanisms are shared in cancer and pregnancy. Through cross-analyzing single-cell RNA-sequencing data from multiple human cancer types and the maternal-fetal interface, we found B7-H4 (VTCN1) is an onco-fetal immune tolerance checkpoint. We showed that genetic deficiency of B7-H4 resulted in immune activation and fetal resorption in allogeneic pregnancy models. Analogously, B7-H4 contributed to MPA/DMBA-induced breast cancer progression, accompanied by CD8+ T cell exhaustion. Female hormone screening revealed that progesterone stimulated B7-H4 expression in placental and breast cancer cells. Mechanistically, progesterone receptor (PR) bound to a newly identified -58 kb enhancer, thereby mediating B7-H4 transcription via the PR-P300-BRD4 axis. PR antagonist or BRD4 degrader potentiated immunotherapy in a murine B7-H4+ breast cancer model. Thus, our work unravels a mechanistic and biological connection of a female sex hormone (progesterone) to onco-fetal immune tolerance via B7-H4 and suggests that the PR-P300-BRD4 axis is targetable for treating B7-H4+ cancer.

The immune microenvironment of lung adenocarcinoma featured with ground-glass nodules.

Early-stage lung cancer is now more commonly identified in the form of ground-glass nodules (GGNs). Presently, the treatment of lung cancer with GGNs mainly depends on surgery; however, issues still exist such as overtreatment and delayed treatment due to the nonuniform standard of follow-up. Therefore, the discovery of a noninvasive treatment could expand the treatment repertoire of ground-glass nodular lung cancer and benefit the prognosis of patients. Immunotherapy has recently emerged as a new promising approach in the field of lung cancer treatment. Thus, this study presents a comprehensive review of the immune microenvironment of lung cancer with GGNs and describes the functions and characteristics of various immune cells involved, aiming to provide guidance for the clinical identification of novel immunotherapeutic targets.

Polypharmacology of ambroxol in the treatment of COVID-19.

The pandemic of coronavirus disease 2019 (COVID-19) by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still underway. Due to the growing development of severe symptoms, it is necessary to promote effective therapies. Ambroxol [2-amino-3,5-dibromo-N-(trans-4-hydroxycyclohexyl) benzylamine] has long been used as one of the over-the-counter mucolytic agents to treat various respiratory diseases. Therefore, we focused on the mechanism of action of ambroxol in COVID-19 treatment. In vitro and in silico screening revealed that ambroxol may impede cell entry of SARS-CoV-2 by binding to neuropilin-1. Ambroxol could also interact with multiple inflammatory factors and signaling pathways, especially nuclear factor kappa B (NF-κB), to interfere cytokines cascade activated by SARS-CoV-2 internalization. Furthermore, multipathways and proteins, such as the cell cycle and matrix metalloproteinases (MMPs), were identified as significant ambroxol-targeting pathways or molecules in PBMC and lung of severe COVID-19 patients by bioinformatics analysis. Collectively, these results suggested that ambroxol may serve as a promising therapeutic candidate for the treatment of severe SARS-CoV-2 infection.

Lung adenocarcinoma manifesting as subsolid nodule potentially represents tumour in the equilibrium phase of immunoediting.

Lung adenocarcinomas manifesting as subsolid nodules (SSN-LUADs) possess distinct dormant behaviour. This study was designed to compare the immune landscapes of normal lungs (nLungs), SSN-LUADs and LUADs manifesting as solid nodules (SN-LUADs) so as to better understand the status of anti-tumour immunity in SSN-LUADs. Mass cytometry by time-of-flight analysis was performed on 299, 570 single cells from nLung, SSN-LUAD and SN-LUAD tissues. The immune cells were identified by phenotype, and the percentages of different immune cell subclusters were compared between SSN-LUADs, SN-LUADs and nLungs. Elevated percentage of CD8+ T cells were identified in SSN-LUADs compared with in nLungs and SN-LUADs. Elevated CD56bright NK cells and decreased CD56dim NK cells were identified in both SSN-LUADs and SN-LUADs compared with in nLungs. The immune landscape of SSN-LUAD fits the theory of equilibrium phase of immunoediting, thus functional adaptive anti-tumour immunity but impaired innate anti-tumour immunity potentially contributes to the maintaining of its dormant behaviour.

Stepwise evolutionary genomics of early-stage lung adenocarcinoma manifesting as pure, heterogeneous and part-solid ground-glass nodules.

BACKGROUND: This study was designed to unravel the genomic landscape and evolution of early-stage subsolid lung adenocarcinomas (SSN-LUADs) manifesting as pure ground-glass nodules (pGGNs), heterogeneous ground-glass nodules (HGGNs) and part-solid nodules (PSNs). METHODS: Samples subjected to either broad-panel next-generation sequencing (NGS) or whole-exome sequencing (WES) were included. Clinicopathologic and genomic features were compared among pGGN, HGGN and PSN, while tumour evolutionary trajectories and mutational signatures were evaluated in the entire cohort. RESULTS: In total, 247 SSN-LUAD samples subjected to broad-panel NGS and 125 to WES were identified. Compared with PSNs, HGGNs had significantly lower tumour mutation count (P < 0.001), genomic alteration count (P < 0.001), and intra-tumour heterogeneity (P = 0.005). Statistically significant upward trends were observed in alterations involving driver mutations and oncogenic pathways from pGGNs to HGGNs to PSNs. EGFR mutation was proved to be a key early event in the progression of SSN-LUADs, with subsequently two evolutionary trajectories involving either RBM10 or TP53 mutation in the cancer-evolution models. CONCLUSIONS: This study provided evidence for unravelling the previously unknown genomic underpinnings associated with SSN-LUAD evolution from pGGN to HGGN to PSN, proving that HGGN was an intermediate SSN form between pGGN and PSN genetically.

EGFR mutation is not a prognostic factor for CNS metastasis in curatively resected lung adenocarcinoma patients.

INTRODUCTION: For NSCLC patients with complete resection, the prognostic role of EGFR mutation for recurrence, especially for CNS metastasis, is still controversial. In this study, we aimed to identify the characteristics of the recurrence pattern of lung adenocarcinoma based on EGFR mutation status. METHODS: Overall, 888 patients with completely surgically resected LUAD who underwent EGFR mutation status analysis from two Chinese institutions were included. Sites and data of initial recurrence were recorded. The recurrence patterns according to EGFR mutation status were estimated by Kaplan-Meier analysis, and hazard rate curves were generated. RESULTS: 245 (27.6%) of 888 patients suffered from recurrence. Before and after PSM, there were no statistically significant differences between the EGFR mutation and EGFR WT groups for all types of recurrence, including CNS metastasis. Multivariable Cox analysis revealed that EGFR status was not a risk factor for all types of recurrence, including CNS metastasis (HR 0.88, 95% CI 0.54-1.46, p = 0.632). The CNS metastasis hazard curve in the EGFR mutation group showed that the first peak occurred at approximately 24-26 months after surgery, which was 10 months later than that in the EGFR WT group. In addition, the second peak time in the EGFR mutation group was approximately 2 years later than that in the EGFR WT group. CONCLUSIONS: EGFR mutation was not an independent prognostic factor for postoperative recurrence. EGFR-mutated LUADs did not have a clinical course with a higher incidence of CNS metastasis. However, the peak hazards for recurrence of CNS metastasis occur at a later time point in the EGFR mutant group compared with the EGFR wild type group.

Skeletal muscle wasting during neoadjuvant therapy as a prognosticator in patients with esophageal and esophagogastric junction cancer: A systematic review and meta-analysis.

BACKGROUND: Considerable controversies exist regarding the severity of skeletal muscle wasting (SMW) during neoadjuvant therapy (NAT) and its impact on therapeutic outcomes in patients with esophageal or esophagogastric junction cancer (EC/EGJC). This systematic review and meta-analysis aimed to resolve these issues. Particularly, the prognostic value of SMW during NAT was compared to pre-NAT and pre-surgery sarcopenia status. METHODS: We searched PubMed, Embase, and Cochrane Library databases through October 13th, 2021 to identify cohort studies focusing on SMW during NAT and therapeutic outcomes in EC/EGJC patients. Both neoadjuvant chemotherapy and neoadjuvant chemoradiotherapy were studied. A meta-analysis was conducted to quantify SMW and increased sarcopenia during NAT. Therapeutic outcomes include perioperative morbidities and survival profiles. A separate meta-analysis investigating the impacts of pre-NAT/pre-surgery sarcopenia on therapeutic outcomes was synchronously performed. RESULTS: Twenty-five studies with 2706 participants were included in this review. The pooled SMW during NAT were -2.47 cm2/m2 in skeletal muscle index and -0.23 cm2/m2 in psoas muscle index, with wasting proportion reaching 4.44%. The pooled prevalence rate of sarcopenia increased from 53.1% before NAT to 65.8% before surgery. Neoadjuvant chemoradiotherapy, advanced age, and being male were identified as risk factors for severe SMW during NAT. Notably, severe SMW during NAT showed a greater hazard ratio (HR) than pre-NAT and pre-surgery sarcopenia in predicting overall survival (HR 1.92, P < 0.001; HR 1.17, P = 0.036; and HR 1.28, P = 0.011, respectively) and recurrence-free survival (HR 1.51, P = 0.002; HR 1.27, P = 0.008; and HR 1.38, P = 0.006, respectively). However, severe SMW during NAT was not significantly associated with perioperative morbidities. CONCLUSIONS: SMW during NAT is a novel prognosticator that is different from sarcopenia for poor survival in EC/EGJC patients. Interventions aiming at maintaining skeletal muscle during NAT are anticipated to promote therapeutic outcomes.

Spread through air spaces is a common phenomenon of pulmonary metastasized tumours regardless of origins.

OBJECTIVES: Spread through air spaces (STAS) is a unique pattern of invasion in primary lung cancers. However, little is known about STAS in pulmonary metastases (PMs). This study was to investigate the incidence of STAS among PMs and the association between STAS and clinicopathological characteristics of PMs. METHODS: A total of 127 patients who underwent metastasectomy at our institution from June 2009 to December 2019 were retrospectively analysed. Survival analysis was performed in 40 patients with PM from colorectal cancer (CRC). RESULTS: STAS was identified in 33.1% of patients (42 of 127) with PMs. STAS was found in PMs of various primary cancers, including CRC, breast cancer, renal cell carcinoma, cholangiocarcinoma and osteogenic and soft tissue sarcoma, but the incidence varies. PMs originating from epithelial tissue showed higher incidence of STAS than those from mesenchymal tissue (45% vs 11%, P < 0.001). Elder age (P = 0.006) and primary sites (P < 0.001) were significantly correlated with STAS. In patients with PMs from CRC, the presence of STAS was an independent predictor of shorter recurrence-free survival (hazard ratio = 10.25, P = 0.002) and poor overall survival (hazard ratio = 4.75, P = 0.047) by multivariable analysis. CONCLUSIONS: STAS might be a lung-specific tumour invasion pattern and STAS is commonly observed in PMs of different origins. The incidence of STAS was significantly higher in PMs originating from epithelial tissues than those from mesenchymal tissues. Presence of STAS was an independent predictor of poor prognosis in patients with PM from CRC.

MicroRNA-98-5p modulates cervical cancer progression via controlling PI3K/AKT pathway.

To probe into the potential mechanism of microRNA (miR)-98-5p inhibiting the biological progress of cervical cancer (CC) cells via regulating PI3K/Akt pathway. Reverse transcription quantitative polymerase chain reaction was applied to detect miR-98-5p expression in CC tissues and cell lines; Cell counting kit-8 and Edu analysis were performed for checking cell proliferation, flow cytometry for cell apoptosis, transwell for cell invasion and migration, Western blot for proliferation-related proteins Ki67 and Proliferating cell nuclear antigen expression, apoptosis-related proteins Bcl-2 and Bax expression, epithelial-mesenchymal transition (EMT)-related proteins Snail, matrix metalloproteinase-3, E-cadherin and N-cadherin expression, as well as PI3K/Akt pathway-related proteins PTEN, PI3K as well as Akt expression levels, and the nude mouse tumor xenograft experiment was applied to verify in vivo. The result clarified, miR-98-5p was reduced in CC. Overexpression miR-98-5p could inhibit CC cell proliferation, invasion, migration and EMT, whereas promoted its apoptosis, but silencing miR-98-5p was opposite. Overexpression miR-98-5p could depress the activation of PI3K/Akt pathway in CC in vivo and in vitro. MiR-98-5p targeted CBX5. In short, miR-98-5p is able to be used as a potential target for treating CC in future research.

MiR-101-3p Suppresses Progression of Cervical Squamous Cell Carcinoma by Targeting and Down-Regulating KPNA2.

Objective We explored mechanism of microRNA-101-3p/Karyopherin α2 (KPNA2) axis in cervical squamous cell carcinoma. Methods: Bioinformatics methods were applied to identify genes for the study. Cell functional assays were implemented to examine the role of the genes in malignant progression of cervical squamous cell carcinoma. Targeting relationship between genes was verified by dual-luciferase assay. Results: MicroRNA-101-3p was lowly expressed in cervical squamous cell carcinoma, while KPNA2 was highly expressed. Dual-luciferase assay identified direct targeting relationship between microRNA-101-3p and KPNA2. Functional assays manifested that highly expressed microRNA-101-3p suppressed cervical squamous cell carcinoma cell growth by targeting KPNA2. Conclusion: Overall, microRNA-101-3p/KPNA2 axis can play an important part in progression of cervical squamous cell carcinoma.

A cross-sectional study of psychological burden in Chinese patients with pulmonary nodules: Prevalence and impact on the management of nodules.

BACKGROUND: Uncertainty after the detection of pulmonary nodules (PNs) can cause psychological burden. We designed this study to quantitatively evaluate the prevalence, severity and possible impact of this burden on the preference of patients for management of nodules. METHODS: The Hospital Anxiety and Depression Scale (HADS) was used to evaluate psychological burden in patients. An independent t-test and a Mann-Whitney U test were used to determine the significance of differences between groups in continuous variables. A chi-square test was used to determine the significance of difference between groups in categorical variables. RESULTS: A total of 334 inpatients diagnosed with PNs were included in the study. A total of 17.96% of the participates screened positive for anxiety and 14.67% for depression. Female patients had significantly higher positive rates of both anxiety and depression screenings than male patients (21.57% vs. 12.31%, p = 0.032 and 18.05% vs. 9.30%, p = 0.028, respectively). Among patients screened positive for anxiety, the proportion of those who chose more aggressive management was significantly higher (34/60 vs. 113/274, p = 0.029). The rate of benign or precursor disease resected was significantly higher in patients with more aggressive management (46.94% vs. 9.63%, p < 0.01). CONCLUSIONS: Anxiety and depression are common in Chinese patients with PNs. Patients with positive HADS anxiety screening results are more likely to adopt more aggressive management that leads to a higher rate of benign or precursor disease resected/biopsied. This study alerts clinicians to the need to assess and possibly treat emotional responses.

Propensity-matched analysis of adjuvant chemotherapy for completely resected Stage IB non-small-cell lung cancer patients.

BACKGROUND: The use of adjuvant chemotherapy (ACT) in completely resected stage IB non-small cell lung cancer (NSCLC) is still controversial. The divergent outcomes of prospective trials have created uncertainty as to the utility of ACT in stage IB NSCLC. This study assesses the effect of postoperative adjuvant chemotherapy in stage IB patients in clinical practice. METHODS: Patients with pT2aN0M0 stage IB NSCLC who underwent complete resection from 2004 to 2015 were identified from prospectively collected databases in two medical centers. The log-rank test was used to compare overall survival (OS) and disease free survival (DFS). Fine and Gray's competing risks regression model was built to identify predictors of cancer-specific survival. One to one propensity-score matching (PSM) was performed to reduce the selection bias and additional analyses were performed on these subgroups. RESULTS: Of 1005 patients identified for the study, 202 (20.1%) received ACT and 803 (79.9%) underwent surgery alone (observation group). Compared with the observation group, patients who underwent ACT were younger (p < 0.001), had larger tumors (p = 0.004), and had higher rates of squamous cell carcinoma (p < 0.001) and lymphovascular invasion (p = 0.017). After propensity score matching, 196 pairs of patients were 1:1 matched in the two groups and all baseline characteristics were well balanced. ACT was not associated with improved survival (including OS, DFS; all log-rank p > 0.05) in both unmatched and matched (196 pairs) cohorts. In subgroup analysis of the matched population, ACT was not associated with survival benefits for patients regardless of whether their tumors measured <4 cm or ≥4 cm (both log-rank p > 0.05). CONCLUSIONS: In patients with completely resected stage IB (T2aN0M0) NSCLC, ACT is not associated with improved prognosis. Further large multicenter studies are needed to confirm these findings.

The value of mediastinoscopy in N staging of clinical N2 lung cancer.

BACKGROUND: To determine the value of mediastinoscopy in N staging of lung cancer with clinical N2 disease. METHODS: We retrospectively reviewed 87 patients who received mediastinoscopy for known or suspected lung cancer, including 83 cervical mediastinoscopies and 4 parasternal mediastinoscopies. All patients were clinically staged N2 for enlarged ipsilateral mediastinal and/or subcarinal lymph nodes (short axis >1.0 cm) on computed tomography scan. RESULTS: Of the 87 patients, 61 cases proved to be N2 disease by mediastinoscopy; the other 26 mediastinoscopy-negative patients underwent thoracotomy for lung resection and mediastinal lymph node dissection in the same operation. Final pathologic N staging was consistent with mediastinoscopic sampling and surgical dissection in 24 patients, and N2 disease was found in 2 patients (false-negative by mediastinoscopy). The sensitivity, specificity, and accuracy of mediastinoscopy were 96.8%, 100%, and 97.7%, respectively. Among all 87 mediastinoscopic procedures, there was no mortality and only 1 complication (1.1%). CONCLUSIONS: Mediastinoscopy is a highly effective and safe procedure for the mediastinal staging of lung cancer with clinical N2 disease.

Video-assisted thoracic surgery for thymoma: long-term follow-up results and prognostic factors-single-center experience of 150 cases.

BACKGROUND: The use of video-assisted thoracic surgery (VATS) in the treatment of thymoma is becoming more and more common. The aim of this study was to evaluate safety and effectiveness of thoracoscopic treatment of thymoma and, more importantly, to report long-term follow-up results of a large single-center cohort. METHODS: A retrospective review of a prospective database was performed. In total, 150 patients who underwent VATS resection for thymoma at Peking University People's Hospital from April 2001 to November 2014 were retrospectively reviewed. RESULTS: Average operation time was 140.1±54.2 min, median blood loss was 50 mL (range, 10-700 mL), median post-operative drainage time was 3 days (range, 1-11 days), and median length of post-op stay was 5 days (range, 2-20 days); 134 patients (89.3%) were followed up successfully. Median follow-up was 59.5 months (range, 2-187 months). Five- and 10-year recurrence free survival (RFS) rates of entire group were 96.5% and 94.4%, respectively; 5- and 10-year RFS rates for Masaoka stages I + II were 98.1% and 98.1%, respectively; 5- and 6-year RFS rates for Masaoka stage III were 90% and 60%, respectively. One case of recurrence in five Masaoka stage IV patients was observed, and 4-year RFS was 80%. Multivariable analysis indicated that recurrence tended to occur in Masaoka stages III + IV patients (P=0.037, HR =12.69, 95% CI: 1.17-138.22) and older patients had a lower risk of recurrence (P=0.029, HR =0.87, 95% CI: 0.77-0.99). Myasthenia gravis (MG) presented in 44 patients (29.3%), of which 36 patients (81.8%) were followed up. Nine patients achieved complete remission, and 19 patients had symptom improvement after surgery. Overall response rate of MG was 77.8% (28/36). CONCLUSIONS: VATS was a safe and effective procedure for treatment of thymomas with satisfactory prognosis. MG with thymoma treated by VATS had comparable neurological outcome to that associated with the trans-sternal approach.

Near-infrared fluorescence-guided thoracoscopic surgical intervention for postoperative chylothorax.

OBJECTIVES: Chylothorax is an infrequent but relatively serious complication after lung cancer surgery. Finding the leakage site and identifying the thoracic duct (TD) are the key points of surgical intervention for chylothorax. In this study, for the first time, we demonstrated near-infrared fluorescence imaging with indocyanine green (ICG) in video-assisted thoracoscopic surgery (VATS) intervention for chylothorax in humans. METHODS: This study included 4 patients diagnosed with chylothorax after lung cancer surgery who underwent VATS intervention; 0.2 mg/kg of ICG was injected subcutaneously into the bilateral inguinal region approximately 30 min before surgery. The D-light P® near-infrared thoracoscope was used for intraoperative fluorescence imaging. RESULTS: All patients underwent VATS intervention on the right side. Chyle leakage points were detected at the point of dissection of the station of the 4R lymph node behind the azygos vein in 3 patients and at the adjacent point of prophylactic ligation of the TD in 1 patient. The fluorescent hotspot of ICG leakage was detected first. The signal-to-background ratio of the TD or chyle averaged 4.41 (range 2.31-6.72). The TD fluorescent signals lasted for at least 1 h. With the guidance of real-time fluorescence lymphography, the fistulas and the main trunk of the TD were identified and double ligated. Small branches of the TD were occasionally detected and ligated. Chylothorax was managed successfully by surgical interventions in all patients. CONCLUSIONS: Near-infrared fluorescence imaging with ICG provided highly sensitive and real-time imaging of the TD in VATS intervention for chylothorax in humans. Clinical registration number: NCT02611245.

Role of Berberine in the Treatment of Methicillin-Resistant Staphylococcus aureus Infections.

Berberine is an isoquinoline alkaloid widely used in the treatment of microbial infections. Recent studies have shown that berberine can enhance the inhibitory efficacy of antibiotics against clinical multi-drug resistant isolates of methicillin-resistant Staphylococcus aureus (MRSA). However, the underlying mechanisms are poorly understood. Here, we demonstrated that sub-minimum inhibitory concentrations (MICs) of berberine exhibited no bactericidal activity against MRSA, but affected MRSA biofilm development in a dose dependent manner within the concentration ranging from 1 to 64 µg/mL. Further study indicated that berberine inhibited MRSA amyloid fibrils formation, which consist of phenol-soluble modulins (PSMs). Molecular dynamics simulation revealed that berberine could bind with the phenyl ring of Phe19 in PSMα2 through hydrophobic interaction. Collectively, berberine can inhibit MRSA biofilm formation via affecting PSMs' aggregation into amyloid fibrils, and thereby enhance bactericidal activity of antibiotics. These findings will provide new insights into the multiple pharmacological properties of berberine in the treatment of microbial-generated amyloid involved diseases.

[Anaerobic biodegradation pathway of the main ingredient of traditional Chinese medicine wastewater].

A laboratory-scale experiment was done. And the baicalin anaerobic biodegradation curve and pathway were discussed by analyzing the results of GC-MS. After analogue of status's trend and analysis of its regression, the biodegradation kinetic equation of baicalin was obtained. And the possible biodegradation pathway was "desugar --> debentures --> open benzene ring --> saturating --> disconnect links". It took at least 47 h to biodegrade baicalin. The main products of anaerobic biodegradation were micro-molecule oxide, such as alcohols, aldehydes, acids, esters, olefin, and hydrocarbon of methane series. This result demonstrated that the rate-limiting steps of bacalin biodegradation were the hydrolyzing and acidification phase. It was suitable to use two-phase anaerobic process and long HRT to biodegrade baicalin.