Recent Advances in the Application of 3D-Printing Bioinks Based on Decellularized Extracellular Matrix in Tissue Engineering.

In recent years, 3D bioprinting with various types of bioinks has been widely used in tissue engineering to fabricate human tissues and organs with appropriate biological functions. Decellularized extracellular matrix (dECM) is an excellent bioink candidate because it is enriched with a variety of bioactive proteins and bioactive factors and can provide a suitable environment for tissue repair or tissue regeneration while reducing the likelihood of severe immune rejection. In this Review, we systematically review recent advances in 3D bioprinting and decellularization technologies and comprehensively detail the latest research and applications of dECM as a bioink for tissue engineering in various systems, with the aim of providing a reference for researchers in tissue engineering to better understand the properties of dECM bioinks.

SIRPG promotes lung squamous cell carcinoma pathogenesis via M1 macrophages: a multi-omics study integrating data and Mendelian randomization.

Background: Squamous cell carcinoma of the lung (LUSC) is a severe and highly lethal malignant tumor of the respiratory system, and its molecular mechanisms at the molecular level remain unc\lear. Methods: We acquired RNA-seq data from 8 surgical samples obtained from early-stage LUSC and adjacent non-cancerous tissues from 3 different centers. Utilizing Deseq2, we identified 1088 differentially expressed genes with |LogFC| > 1 and a p-value < 0.05 threshold. Furthermore, through MR analysis of Exposure Data for 26,153 Genes and 63,053 LUSC Patients, incorporating 7,838,805 SNPs as endpoints, we identified 213 genes as potential exposure factors. Results: After intersecting the results, we identified 5 differentially expressed genes, including GYPE, PODXL2, RNF182, SIRPG, and WNT7A. PODXL2 (OR 95% CI, 1.169 (1.040 to 1.313)) was identified as an exposed risk factor, with p-values less than 0.01 under the inverse variance weighted model. GO and KEGG analyses revealed enhanced ubiquitin-protein transferase activity and activation of pathways such as the mTOR signaling pathway and Wnt signaling pathway. Immune infiltration analysis showed downregulation of Plasma cells, T cells regulatory (Tregs), and Dendritic cells activated by the identified gene set, while an enhancement was observed in Macrophages M1. Furthermore, we externally validated the expression levels of these five genes using RNA-seq data from TCGA database and 11 GEO datasets of LUSC, and the results showed SIRPG could induce LUSC. Conclusion: SIRPG emerged as a noteworthy exposure risk factor for LUSC. Immune infiltration analysis highlighted Macrophages M1 and mTOR signaling pathway play an important role in LUSC.

Clinical application of preserving spontaneous breathing non-intubation anesthesia in thoracoscopic surgery for lung cancer under ERAS concept.

OBJECTIVE: To explore the safety and feasibility of thoracoscopic surgery in patients with lung cancer under non-intubation anesthesia, and to evaluate the advantages of the non-intubation anesthesia compared with intubation anesthesia on enhanced recovery after surgery (ERAS). METHODS: A retrospective cohort study was conducted in which 100 patients who underwent thoracoscopic lung cancer surgery from January 2020 to February 2021 in the Department of Thoracic Surgery of the First Affiliated Hospital of Soochow University were included and divided into non-intubation group (n = 50) and intubation group (n = 50). The primary outcome was the comparison of intra- and postoperative parameters. Secondary outcomes included inflammatory response indicators and intra- and postoperative complications. RESULTS: There was no significant difference between the two groups in anesthesia effect score, blood loss, lowest pulse oxygen saturation, operation time, postoperative chest tube indwelling time (P > 0.05). Non-intubation group had less intraoperative remifentanil dosage, less change of blood pressure and heart rate, lower postoperative pain numerical score(NRS), less medical costs, smaller incidence rate of throat discomfort (P < 0.05). The non-intubation group was also associated with less extubation time, postanesthesia care unit recovery time, ambulation time, food intake time, postoperative antibiotic use time, and hospital stay (P < 0.05). The increase of C-reactive protein in the non-intubation group was lower than that in the intubation group (P < 0.05). CONCLUSION: Non-intubation anesthesia for thoracoscopic lung cancer surgery is safe and feasible. Compared with the intubation anesthesia, it has advantages in ERAS and reducing medical costs.

The role of seven autoantibodies in lung cancer diagnosis.

BACKGROUND: To investigate the expression and diagnostic value of seven autoantibodies (P53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGE, and CACE) in lung cancer patients. METHODS: A total of 370 patients were admitted to the Thoracic Surgery of the First Affiliated Hospital of Suzhou University from 2017 to 2019, including 305 patients with lung cancer and 65 patients with benign lesions. The concentrations of the seven autoantibodies were determined by enzyme linked immunosorbent assay (ELISA).The expression levels of each antibody were compared between the two groups, and the levels of each antibody between lung cancer patients with different pathological types were also compared. We aimed to analyze the diagnostic efficiency of single antibody detection combined with seven antibodies, and also to explore whether there were differences among the positive rates of each antibody in sex, age, smoking history, pathological classification, and clinical stages in the lung cancer group. RESULTS: The expression levels of seven autoantibodies in the lung cancer group were higher than those in the benign lesion group. In the lung cancer group, the expression levels of the seven autoantibodies did not vary statistically among different pathological types. The area under the curve of combined detection of the seven antibodies reached 0.735, and the Y-index reached 0.35, which was higher than that of single antibody detection. P53 exhibited the highest sensitivity and lowest specificity; meanwhile, PGP9.5, SOX2, GAGE7, GBU4-5, and MAGEA1 exhibited low sensitivity and high specificity. The sensitivity and specificity of the CAGE were approximately 60%, respectively. There was no statistical difference in the positive rate of each antibody in age, smoking history, and clinical stage. The positive rate of MAGEA1 and CAGE was statistically different in sex, and the positive rate of MAGEA1 was statistically different in pathological classification. CONCLUSIONS: The seven autoantibodies of lung cancer can potentially be used as an auxiliary examination method for the early diagnosis of lung cancer.

RPB5-mediating protein promotes the progression of non-small cell lung cancer by regulating the proliferation and invasion.

BACKGROUND: This study aimed to investigate the relationship between RNA polymerase II subunit 5 (RPB5)-mediating protein (RMP) and clinicopathological characteristics of non-small cell lung cancer (NSCLC) patients by measuring the expression level of RMP in human NSCLC tissues and cell lines. At the same time, we studied the impact of RMP on the biological function of cancer, providing strong support for gene targeted therapy of NSCLC. METHODS: Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were used to determine the expression levels of messenger (m)RNA and protein in NSCLC cell lines and tissues. Cell counting kit 8 (CCK8) assay and flow cytometry were selected to detect cell proliferation, cycle and apoptosis. The wound healing assay was chosen to detect the migration and invasion ability of cells. The xenograft model was performed to study the function of RMP in vivo. Immunohistochemical (IHC) staining showed the levels of RMP, Bcl-2, Bax and caspase-3. RESULTS: First, mRNA and protein levels of RMP were relatively overexpressed in NSCLC cells. Compared with the corresponding normal tissues, the mRNA and protein levels of RMP were significantly higher in human NSCLC tissues. Concurrently, we found that the expression of RMP was related to the status of lymph nodes (LNs) in cancer tissues and T stage. Then, RMP overexpression promoted the proliferation of A549. At the same time, RMP provided A549 cells the ability to resist chemotherapy and radiotherapy; when A549 cells were treated with gefitinib and radiation, RMP reduced apoptosis. We also found that RMP can protect A549 from G2 block caused by radiation. Over-irradiated RMP-overexpressed A549 cells had lower Bcl2-associated X protein (Bax) levels and higher B-cell lymphoma 2 (Bcl-2) levels. The migration and invasion ability of A549 cells was increased by RMP. Finally, RMP can promote tumor growth by increasing Bcl-2 levels and decreasing Bax and caspase-3 levels in the xenograft model. CONCLUSIONS: There is potential for RMP to develop into a diagnostic and therapeutic target for NSCLC.

The Diagnosis of Intrapulmonary Metastasis Multifocal Pulmonary Ground-Glass Nodules Based on Oncogenic Driver Mutation: Two Case Reports and Review of Literature.

With the increased use of low-dose computed tomography (LDCT), the detection of multifocal pulmonary ground-glass nodules (GGNs) has increased. According to the current clinical guidelines, multifocal GGNs tend to be treated as the multiple primary early-stage lung adenocarcinoma. However, studies have indicated that parts of multiple GGNs may originate from single nodules via intrapulmonary metastasis (IPM). Such IPM indicates the advanced stages even when the multiple GGNs are present as the early characteristics in pathological assessments. However, no gold standard exists for the differential diagnosis of multiple IPM GGNs. Here, we report two multifocal pulmonary GGNs cases where panel sequencing (672 driver mutation loci) showed that patient 1 (P1) shared two rare epidermal growth factor receptor (EGFR) mutations (primary L747_T751del and primary T790M) in the left upper lobe anterior segment and left lower lobe superior segment, respectively. Patient 2 (P2) shared a low-frequency human epidermal growth factor receptor 2 (HER2) mutation (primary Tyr772_Ala775dup) in two GGNs located in the apicoposterior and superior lingular segment of the left lower lobe (LLL). Oncogenic driver mutations were concordant between primary tumors and metastasis. Thus, shared low-frequency driver mutations in multiple GGNs strongly suggested that IPM existed with a high probability in these patients. Also, tumor cell spread through air spaces (STAS) was identified in pathological sections of the left upper lobe (LUL) nodule of P1, suggesting aerogenous spread may have been an effective pathway for IPM. Our report suggests that oncogenic driver mutations have prominent diagnostic value for IPM. Also, GGN IPM may occur in one lung lobe and between in different lung lobes.

Bronchoplasty for treating the whole lung atelectasis caused by endobronchial tuberculosis in main bronchus.

BACKGROUND: Patients with the whole lung atelectasis caused by endobronchial tuberculosis (EBTB) in main bronchus usually undergo pneumonectomy, which may result in a large number of complications and poor quality of life. In the study, we summarized our experience with bronchoplasty treatment for the whole lung atelectasis caused by EBTB in main bronchus, in which long-term medical therapy failed. METHODS: Eight patients with whole lung atelectasis, who suffered from EBTB in main bronchus, were treated by bronchoplasty: 3 patients were treated by bronchoplasty for the left main bronchus; 2 patients were treated by sleeve resection of right upper and middle lobes; and 3 patients were treated by sleeve resection of left upper lobe. The patients were followed up for a period of time. RESULTS: All the patients were cured by surgery, were discharged from hospital uneventfully, had anastomotic patency and good lung expansion. Neither bronchopleural fistula nor recurrence was observed. CONCLUSIONS: Bronchoplasty is an effective treatment for atelectasis of the whole lung caused by EBTB in the main bronchus, in which long-term medical therapy failed. Prognosis was favorable.

Effect of FAM196B in human lung adenocarcinoma.

Lung cancer is the leading cause of cancer-related mortality worldwide. The present study focused on the function of family with sequence similarity 196 member B (FAM196B) in lung adenocarcinoma (LUAD). We analyzed lung carcinoma patients' data from the Cancer Genome Atlas (TCGA), and verify the change of FAM196B expression in 30 LUAD tissues and matched adjacent non-tumor tissues (> 5 cm) by tissue microarray. To investigate the role of FAM196B in LUAD, we used lentivirus-mediated short hairpin RNA (shRNA) to knockdown FAM196B expression in human LUAD cell line A549 and H1299 and assessed it by RT-qPCR and western blot. Celigo Imaging Cytometry System, MTT assays and colony formation were conducted to evaluate cell proliferation. Cell apoptosis were assayed by Annexin V staining. We found that FAM196B was significantly upregulated (P=5.06E-06) in LUAD compared with adjacent non-tumor tissues. Cell proliferation was inhibited in FAM196B-silenced A549 and H1299 cells using Celigo Imaging Cytometry System, MTT assays and colony formation assays. Apoptosis rate was significantly increased in FAM196B-shRNA group than the control group. In conclusion, Knockdown of FAM196B can inhibit cell proliferation, cell colony formation capacity, and promote cell apoptosis in A549 and H1299 cell lines. FAM196B may be one novel potential targets for treating patients with LUAD.

The Expression and Clinical Signification of PD-1 in Lymph Nodes of Patients with Non-small Cell Lung Cancer.

To screen anti-programmed cell death protein 1 (PD-1) antibody treatment of the dominant population, it is necessary to understand the expression of PD-1 in tumor metastasis microenvironment. The aim of the present study was to detect the expression of PD-1 in lymph nodes of 51 patients with non-small cell lung cancer (NSCLC) by using flow cytometry (FCM). The results showed that the PD-1 expression on CD3+ T cells was significantly increased in NSCLC metastatic lymph nodes (50.08 ± 8.03%) compared with nonmetastatic lymph nodes (36.25 ± 11.27%) (t = 5.208, p < 0.001).We also found that PD-1 expression was not associated with age, sex, and smoking, and it is associated with pathological type and staging of lung cancer. This study demonstrated that PD-1 may involve in lymph nodes metastasis and promote the understanding of the mechanism of immunotherapies in the NSCLC.

The PDRG1 is an oncogene in lung cancer cells, promoting radioresistance via the ATM-P53 signaling pathway.

PDRG1, is short for P53 and DNA damage-regulated gene, which have been found over 10 years. Although severe studies have described the roles of PDRG1 separately in many kinds of tumors, how to act as an oncogene are unclear. To better verify the function of PDRG1 in lung cancer, both loss-function and gain-function of PDRG1 studies based on two human lung cancer lines were performed. Following the transfection of PDRG1, both A549 and 95-D cells showed significant changes in cell viability, the expression of some protein and apoptosis, which were all implied the PDRG1 is an oncogene. Another interesting finding is PDRG1 could promote radioresistance involved the ATM-p53 signaling pathway in lung cancer. If we combine radiotherapy with gene-targeted therapy together effectively, predominant effect may be acquired, which is a huge milestone in clinical cure about lung cancer.