Amelioration of ligamentum flavum hypertrophy using umbilical cord mesenchymal stromal cell-derived extracellular vesicles.

Ligamentum flavum (LF) hypertrophy (LFH) has been recognised as one of the key contributors to lumbar spinal stenosis. Currently, no effective methods are available to ameliorate this hypertrophy. In this study, human umbilical cord mesenchymal stromal cell-derived extracellular vesicles (hUCMSC-EVs) were introduced for the first time as promising vehicles for drug delivery to treat LFH. The downregulation of miR-146a-5p and miR-221-3p expressions in human LF tissues negatively correlated with increased LF thickness. The hUCMSC-EVs enriched with these two miRNAs significantly suppressed LFH in vivo and notably ameliorated the progression of transforming growth factor ß1(TGF-ß1)-induced fibrosis in vitro after delivering these two miRNAs to mouse LF cells. The results further demonstrated that miR-146a-5p and miR-221-3p directly bonded to the 3'-UTR regions of SMAD4 mRNA, thereby inhibiting the TGF-ß/SMAD4 signalling pathway. Therefore, this translational study determined the effectiveness of a hUCMSC-EVs-based approach for the treatment of LFH and revealed the critical target of miR-146a-5p and miR-221-3p. Our findings provide new insights into promising therapeutics using a hUCMSC-EVs-based delivery system for patients with lumbar spinal stenosis.

A prediction model integrated genomic alterations and immune signatures of tumor immune microenvironment for early recurrence of stage I NSCLC after curative resection.

BACKGROUND: Surgery is the standard treatment for patients with stage I non-small cell lung cancer (NSCLC). However, postoperative recurrence leads to a poor prognosis of patients. Currently, there is no effective prognostic biomarker and perioperative treatment for patients with stage I NSCLC. METHODS: One hundred thirty stage I NSCLC patients who had surgical resection were enrolled, including 69 patients who had recurrence within three years and 61 patients who had no recurrence (follow up more than five years). Whole exome sequencing was performed to evaluate gene mutation, copy number variation, and tumor mutation burden (TMB). Immunohistochemistry was carried out to assess the expression of PD-L1 and the level of CD3+ and CD8+ tumor-infiltrating lymphocytes (TILs). Tumor mutation score (TMS) was constructed with the recurrence-associated genes identified by Lasso regression. Immunoscore (IS) was built based on the location and density of CD3+ and CD8+ TILs. Logistic regression was performed to build a prediction model. Seventy percent of patients were included in the training cohort and 30% patients in the testing cohort. P<0.05 was considered to be statistically significant. RESULTS: Univariate analysis showed that lung adenocarcinoma (LUAD), MUC4 mutation, and high TMB were related to early recurrence (P=0.008, 0.0008, and <0.0001, respectively). CD3+ and CD8+ TILs within tumor center and invasive margin significantly negatively correlated with recurrence. EGFR mutation and PD-L1 expression had no association with recurrence. Early recurrence group had significantly higher TMS and lower IS (P<0.0001 and P=0.0003, respectively). Multivariate analysis showed that high TMS and low IS were independent predictors for early recurrence (P<0.0001 and P=0.001, respectively). Integrating TMS and IS, we built a recurrence-model with great discrimination and calibration in the training cohort (AUC =0.935; HL test, P=0.2885) and testing cohort (AUC =0.932; HL test, P=0.5515). CONCLUSIONS: High TMS and low IS were both poor prognostic factors for recurrence in stage I NSCLC. The integrated recurrence-model helps identify patients with high recurrence risk, which provides evidence for future research about perioperative treatment.

Surgical Outcomes After Neoadjuvant Chemoimmunotherapy for Resectable Non-Small Cell Lung Cancer.

BACKGROUND: Neoadjuvant chemoimmunotherapy for resectable non-small cell lung cancer (NSCLC) represents an important research topic. Despite the potential benefits of this approach, the inflammatory responses and adverse events associated with neoadjuvant chemoimmunotherapy can present technical challenges and compromise a planned resection. This study assessed the safety and feasibility of neoadjuvant chemoimmunotherapy followed by surgery for resectable NSCLC. METHODS: The study was conducted from May 2019 to March 2021. Patients who were age 18 years or older, were diagnosed with stage Ib-IIIb NSCLC, and received neoadjuvant chemoimmunotherapy followed by surgery were included. Demographic information, clinical and pathologic characteristics, data about neoadjuvant therapy, and surgical details were collected by retrospective chart review. Toxicity profiles were collected retrospectively or by telephone follow-up. RESULTS: Twenty patients were included in this study. The median age was 56 years (range, 48-72 years), and 18 patients (90%) were men. Squamous carcinoma (14/20, 70%) was the most common cancer type, followed by adenocarcinoma (4/20, 20%), adenosquamous carcinoma (1/20, 5%), and large cell neuroendocrine carcinoma (1/20, 5%). All patients received two to four cycles of neoadjuvant therapy, and the median interval between final therapy and surgery was 49 days (range, 23-133 days). Computed tomography evaluation after neoadjuvant therapy showed partial response in 15 patients (75%) and stable disease in 5 (25%). Final pathologic examinations showed major pathologic response in eight patients, including pathologic complete response in five (25%). Most patients (18/20, 90%) had reduced pathologic staging. Twelve patients (60%) underwent open thoracotomy; the other eight patients underwent minimally invasive surgery, which was uneventful and without intraoperative conversion to open thoracotomy. No perioperative deaths occurred, and only seven patients (35%) developed postoperative complications. Most patients experienced only grade 1-2 adverse effects and laboratory abnormalities during neoadjuvant therapy, and no grade 3 or worse adverse effects or laboratory abnormalities occurred. No patients experienced surgical delays as a result of immune-related adverse events. CONCLUSIONS: Preoperative administration of chemoimmunotherapy for patients with resectable NSCLC was safe and feasible.

The clinical and radiological characteristics of pulmonary cryptococcosis in immunocompetent and immunocompromised patients.

BACKGROUND: We characterized the clinical features, radiographic characteristics, and response to treatment of immunocompetent and immunocompromised patients with pulmonary cryptococcosis (PC). METHODS: We retrospectively reviewed the medical records and radiological profiles of patients diagnosed with PC who received surgical resection between May 2015 and November 2020 in a tertiary referral center. RESULTS: A total of 21 males and 18 females were included in the study. 23 patients were immunocompetent and 20 out of the 39 were asymptomatic. Immunocompetent patients were diagnosed with PC at a younger age than immunocompromised patients (48.9 vs 57.1 years, P = 0.02). Single nodule pattern was the most frequent lesion pattern (33 out of 39, 84.6%) and right upper lobe was the most common site of location (15 out of 47, 31.9%). The majority of lesions were located peripherally (38 out of 47, 80.9%) and most lesions were 1-2 cm in diameter (30 out of 47, 63.8%). Cavitation was more likely to occur in immunocompromised patients (5 out of 11, 45.5%) than in immunocompetent patients (6 out of 36, 16.7%) (P = 0.04) and there was complete resolution of PC in all patients treated with anti-fungal therapy. CONCLUSIONS: Immunocompetent patients were diagnosed with PC at a younger age than immunocompromised patients. Single nodule pattern was the most frequent lesion pattern in PC patients. Cavitation was more likely to occur in immunocompromised patients than in immunocompetent patients.

Roles of the hepcidin-ferroportin axis and iron in cancer.

Hepcidin is a low-molecular-weight hepatic peptide that regulates iron homeostasis, and acts by causing the degradation of its receptor, the cellular iron exporter ferroportin. On the basis of the major role of the hepcidin-ferroportin axis in iron regulation, recently several studies have discussed its expression and influence on the development and prognosis of cancer. Iron plays a pivotal role in homeostasis. However, insights into the mechanisms of normal iron regulation have provided a new perspective on the basic mechanisms, biological rationale, and pathophysiologic implications of changes in iron metabolism in cancer. Besides being a crucial stimulus for cancer, iron dysfunction also causes cancer-related anemia. In this review, we discuss aspects of the hepcidin-ferroportin axis and iron regulation, as well as the inherent connections between them in cancer. We also attempt to consider the possibility in theory of novel targets for further individualized therapy. However, many molecular mechanisms and functions of these regulators remain unclear.

Pulmonary artery pseudoaneurysm caused by a rare vascular tumor: epithelioid hemangioendothelioma.

We present a case of a 58-year-old female with a rare vascular tumor of intermediate malignancy. The initial manifestation was a pseudoaneurysm caused by the rupture of the right pulmonary artery after tumor invasion. The diagnosis of epithelioid hemangioendothelioma was confirmed by the morphologic and immunocytochemical features after surgery. The patient recovered smoothly and there has been no evidence of local recurrence or metastasis during the 2 years of follow-up.

Relationship between the depression status of patients with resectable non-small cell lung cancer and their family members in China.

PURPOSE: Less work on depression status has been done with family members of patients with non-small cell lung cancer (NSCLC). This study investigated depression status of patients and their family members; and the relationship of the depression status between these two groups. METHODS: This cross-sectional study enrolled 194 patients diagnosed with non-small cell lung cancer as well as their family members. In this study, a self-administered General Information Questionnaire was used to collect general information and the Self-rating Depression Scale (SDS) to assess depression status. Linear correlation analysis was used to probe the relationship of the depression status between patients and their family members. RESULTS: Of the 194 patients, 148 (76.3%) showed symptoms of depression. 148 (76.3%) family members had depression symptoms. The severity of depression in patients was positively correlated with that of family members (r = 0.577, p < 0.01). CONCLUSIONS: Patients with lung cancer and their family members suffered depression, and the two were correlated. A prospective study might prove helpful in determining the real relationship existing between the two groups' mental status and whether early detection and intervention might ameliorate this current situation.

Recent advances and prospects in the isolation by size of epithelial tumor cells (ISET) methodology.

Current technologies to identify and characterize circulating tumor cells (CTCs) and _ circulating tumor microemboli (CTMs) among hundreds of millions of leukocytes in the bloodstream can be classified into tumor-marker-dependent and -independent technology. Isolation by size of epithelial tumor cells (ISET) is a tumor-marker-independent technology, in which CTCs are isolated by filtration without use of tumor-associated markers, as a result of their large size relative to circulating blood leukocytes. ISET allows cytomorphological, immunocytological, and genetic characterization of CTCs and CTMs. It offers a number of advantages, including retention of cell morphology; non-antigen dependence; amenability of cells to further interrogation by immunolabeling, fluorescence in situ hybridization, and RNA/DNA analysis; ability to isolate CTMs; reliability. Therefore, morphological-analysis-based and antigen-independent ISET methodology can yield more accurate and objective characterization of epithelial-mesenchymal transition. We can evaluate efficacy of _chemotherapy and radiotherapy and other cancer-targeting therapies by using xenografts that are suitable models for mechanistic studies of ISET-isolated CTC/CTM biology. In addition, a new _ISET-based device could be designed to increase sensitivity to CTCs/CTMs greatly and reduce the number of CTCs/CTMs directly during the blood flow, thus decreasing the _possibility of tumor recurrence and metastasis while retaining normal blood cells. This article reviews recent advances and prospects in ISET methodology and provides new insights into ISET methodology, with important implications for the clinical management of cancer patients.

Comparative proteome analysis of human lung squamous carcinoma using two different methods: two-dimensional gel electrophoresis and iTRAQ analysis.

Discovery of early-diagnosis biomarkers is the key to improve the early-diagnosis and prognosis of human lung squamous carcinoma (hLSC). In order to identify more exhaustive and systematic protein biomarkers for early-diagnosis of hLSC, we chose LCM purifed cells from hLSC tissues and paired normal bronchial epithelia(NBE) tissues and used two methods, the classical 2-DE/MS approach and the new iTRAQ analysis. We found a total of 63 differential proteins, 22 proteins in 2-DE and 59 proteins in iTRAQ analysis, between hLSC and NBE tissues. Among them, 18 proteins were quantified using both methods. The expression level of 15 proteins (68.2%) in 2-DE was consistent with that in iTRAQ analysis. Series of proteins involved in cytoskeleton, chaperone, GTP binding, metabolic process, cell apoptosis, cell proliferation and differentiation, signal transduction, transcription and translation were identified, suggesting their possible role in the emergence of oncogenic pathways leading to carcinogenesis of hLSC. These proteins may make as potential biomarkers for diagnosis of hLSC. The two methods gave us closely related but different information about proteins, suggesting they are complementary or at least supplementary methods at present. Our results show both the usefulness of iTRAQ reagent technology for identification of further potential marker proteins as well as for prevalidation of biomarker.

Identification of candidate biomarkers for early detection of human lung squamous cell cancer by quantitative proteomics.

To discover novel biomarkers for early detection of human lung squamous cell cancer (LSCC) and explore possible mechanisms of LSCC carcinogenesis, iTRAQ-tagging combined with two dimensional liquid chromatography tandem MS analysis was used to identify differentially expressed proteins in human bronchial epithelial carcinogenic process using laser capture microdissection-purified normal bronchial epithelium (NBE), squamous metaplasia (SM), atypical hyperplasia (AH), carcinoma in situ (CIS) and invasive LSCC. As a result, 102 differentially expressed proteins were identified, and three differential proteins (GSTP1, HSPB1 and CKB) showing progressively expressional changes in the carcinogenic process were selectively validated by Western blotting. Immunohistochemistry was performed to detect the expression of the three proteins in an independent set of paraffin-embedded archival specimens including various stage tissues of bronchial epithelial carcinogenesis, and their ability for early detection of LSCC was evaluated by receiver operating characteristic analysis. The results showed that the combination of the three proteins could perfectly discriminate NBE from preneoplastic lesions (SM, AH and CIS) from invasive LSCC, achieving a sensitivity of 96% and a specificity of 92% in discriminating NBE from preneoplatic lesions, a sensitivity of 100% and a specificity of 98% in discriminating NBE from invasive LSCC, and a sensitivity of 92% and a specificity of 91% in discriminating preneoplastic lesions from invasive LSCC, respectively. Furthermore, we knocked down GSTP1 in immortalized human bronchial epithelial cell line 16HBE cells, and then measured their susceptibility to carcinogen benzo(a)pyrene-induced cell transformation. The results showed that GSTP1 knockdown significantly increased the efficiency of benzo(a)pyrene-induced 16HBE cell transformation. The present data first time show that GSTP1, HSPB1 and CKB are novel potential biomarkers for early detection of LSCC, and GSTP1 down-regulation is involved in human bronchial epithelial carcinogenesis.

[Establishment of patient-derived xenotransplantation models for non-small cell lung cancer in immune deficient mice].

BACKGROUND AND OBJECTIVE: Targeted therapies have become a valuable therapeutic option for cancer. Establishment of different animal tumor models has become necessary. This study was to establish xenotransplantation models for patient-derived non-small cell lung cancer (NSCLC) in immune deficient mice. METHODS: Immune deficient mice, BALB/C-nu, NOD/scid and SCID, 16 in each strain, were used. Sixteen tumor specimens were obtained from patients with NSCLC. Each specimen was subcutaneously transplanted into one mouse from each of the three strains. The tumor formation rate, time to tumor engraftment, tumor volume doubling time were recorded and compared among the three strains of mice. Histology of xenograft tumors was examined. RESULTS: The total tumor formation rate was 75% (12/16). The tumor formation rate was the highest in SCID mice (56.25%). Only four tumors were engrafted in SCID mice, and two in BALB/C-nu mice. The tumor formation rate, time to tumor engraftment, and tumor volume doubling time were not significantly different among the three strains of mice. The incidence of tumor size over 1cm in the upper flanks of the mice (56.25%) was significantly higher than that in the lower flanks (25%) (P=0.037). Haematoxylin Eosin staining revealed a high degree of histological similarity between all xenograft and the parental tumors. CONCLUSIONS: We have established xenotransplantation models for patient-derived NSCLC with a success rate of 75% in BALB/C-nu and SCID mice. The xenograft tumors have the same histological features to those of their parental tumors.

[NUSS procedure by video-assisted thoracoscopy for correction of pectus excavatum].

OBJECTIVE: To review the management of NUSS procedure by video-assisted thoracoscopy for the correction of pectus excavatum in 18 patients, and to prevent and treat some complications. METHODS: Eighteen patients with severe pectus excavatum underwent NUSS procedure by video-assisted thoracoscopy from December 2006 to September 2007. RESULTS: The operation time ranged from 30 to 70 min ( average 45 min). Good Results were achieved for all patients after the operation. All of them recovered and were discharged 5 approximately 7 days after the operation. Minor complications occurred in 2 patients. CONCLUSION: NUSS procedure is microinvasive and has beautiful outlook with shorter operation time and simple handling.The minimally invasive technique has a low complication rate with satisfactory short-term result. Proper management is important for the recovery at the early postoperative stage.

Superior vena cava reconstruction using bovine jugular vein conduit.

The glutaraldehyde-treated bovine jugular vein conduit (BJVC) is a xenograft conduit initially used for right ventricular outflow tract reconstruction and has never been used for reconstruction of superior vena cava (SVC). In September 2003, a patient with SVC obstruction underwent SVC reconstruction using BJVC. He has been alive for 42 months and free from signs and symptoms of SVC obstruction except that metastasis was found in the vertebrae. The radionuclide venography showed the graft tube was patent and only slight stenosis was found in the proximal anastomosis. The initial result supports BJVC as an acceptable alternative for SVC reconstruction.

[Reconstructive methods for hypopharynx and cervical esophagus].

OBJECTIVE: To review the surgical treatment for reconstructing hypopharynx and cervical esophagus after hypopharyngo-oesophagectomy, and to evalue its efficacy. METHODS: Different methods were adopted to reconstruct the hypopharynx and cervical esophagus among 25 cases, including 14 cases of carcinoma of the hypopharynx and 11 of carcinoma of hypopharynx and cervical esophagus. In accordance with the standard of the International Union Against Cancer in 1997, the 25 cases were divided into different clinic stages, among which 5 were in T(2)N(0), 2 in T(2)N(1), 4 in T(3)N(0), 3 in T(3)N(1), 7 in T(4)N(1) and 3 in T(4)N(2). Treatment protocol was as follow: Pure operation for 5 cases, re-operation after radiotherapy for 2 cases, operation plus radiotherapy for 18 cases, laryngeal conservation operation for 8, and neck dissection for 21 cases. Reconstruction was done by using free jejunal transplantation, gastric pull-up, the laryngotracheal flap, and myocutaneous flap. RESULTS: After the reconstruction, 3 cases of free jejunal graft and gastric pull-up, 4 of laryngotracheal flap recovered oral fleeding within 2 weeks. No serious complications occurred. After 18 cases underwent the myocutaneous flap reconstruction, no complications occurred in 10 patients, but there were different complications in 8 cases, including pharyngocutaneous fistula (6 cases), haryngoesphageal stenosis (7 cases), and pectoralis major myocutaneous flap necrotic (1 case). The 3-year survival rate was 38.9% (7/18). CONCLUSION: Reconstruction with free jejunal graft, gastric pull-up, and laryngotracheal flap constitutes is a safe and reliable method to restore the continuity of the upper digestive tract after pharyngo-laryngo-oesophagectomy. After the reconstruction with myocutaneous flap, there is high incidence of pharyngocutaneous fistula and haryngoesophageal stenosis.

[Clinical application of video-assisted thoracoscopic surgery].

OBJECTIVE: To explore the clinical application of video-assisted thoracoscopic surgery (VATS). METHODS: We retrospectively analyzed the clinical data of 672 cases of VATS. There were 17 thoracic diseases such as emphysema, bullectomy for spontaneous pneumothorax, massive bullae, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, hydropericardium, malignant pleural fluid, etc. RESULTS: The mean operation time was 57 minutes and there were no intraoperative complications. The bleeding during the operation was less than 100 mL. Postoperative pneumothorax occurred in 4 patients and among them 2 patients were of relapse after 1 month. The intrathoracic drain in most patients was removed with an average of 2. 5 days. A supplementary incision was needed in 10 cases: Six were due to the adhesion of full pleural cavity and 4 were found with the malignant tumor during the operation. CONCLUSION: VATS is an alternative approach that provides a safe, less invasive, and effective operation for treating spontaneous pneumothorax, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, pericardial perfusion, and acute chest trauma patients.

[Modified low-potassium dextran solution in heart-lung transplantation].

OBJECTIVE: To determine the application of modified low-potassium dextran (LPD) solution in heart-lung transplantation. METHODS: We used the modified LPD as the lung flush solution of the donor in the first heart-lung transplantation procedure in Hunan. RESULTS: The patient survived 555 days after the surgery and severe infection, lung disfunction or severe graft rejection never occurred. CONCLUSION: Modified LPD as lung flush solution of the donor during heart-lung transplantation demonstrates an excellent effect of lung protection.

[Expression of PTEN, p16, p21, and p53 proteins in lung cancer using tissue microarray and their clinical significance].

BACKGROUND & OBJECTIVE: The genesis of lung cancer was associated with mutation or abnormal expression of PTEN, p16, p21, and p53. Tissue microarray provides a high throughout tool for genes expression. But little is reported about expression of PTEN, p16, p21, and p53 in lung cancers with tissue microarray. The aim of this study was to investigate the expression of PTEN, p16, p21, and p53 proteins and to analyze their relationship with the pathogenesis, invasion, and metastasis in lung cancer. METHODS: The expression of the antioncogene proteins in 100 cases of lung cancer and corresponding adjacent tissues were determined by tissue microarray combined with immunohistochemistry. RESULTS: The positive expression rates of PTEN, p16, p21, and p53 proteins were 31% (31/100), 38% (38/100), 42% (42/100), 53% (53/100) in lung cancer tissues, and were 85% (85/100), 72% (72/100), 80% (80/100), and 23% (23/100) in the adjacent cancer tissues, respectively, showing a low expression of PTEN, p16, p21 in cancer tissues, and high expression of p53 outside of them (P< 0.05, P< 0.01). Furthermore, the expression of PTEN, P16, and p53 proteins showed positive correlation with the clinical degrees and pathological stages of lung squamous carcinomas and adenocarcinomas (P< 0.05,P< 0.01). In lung cancer with lymph node metastasis, the expression of PTEN, p16, and p21 were low, but the expression of p53 increased significantly (P< 0.05, P< 0.01). CONCLUSION: Tissue microarray provided a useful high-throughout tool for multigene expression in large-scale investigations. There existed low expression of PTEN, p16, p21 proteins and over-expression of mutated p53 protein. Coexpression of these antioncogenes played an important role in invasion and metastasis in lung cancer.