Systemic immune microenvironment and regulatory network analysis in patients with lung adenocarcinoma.

BACKGROUND: This study applied a complex bioinformatics analysis to explore the hub regulators and immune network to further elucidate the molecular mechanisms of lung adenocarcinoma (LUAD) immune regulation. METHODS: LUAD immunological microenvironment features and microenvironment-related differential expression genes (DEGs) were identified by ESTIMATE algorithm and linear models for microarray analyses (LIMMA), respectively. CIBERSORT and Igraph algorithms were applied to construct the LUAD-related immunocyte infiltration and regulatory network. Kaplan-Meier survival analysis, and univariate and multivariate Cox analysis were used to predict independent risk factors and screen for the hub genes. In addition, hub genes-correlated gene set enrichment analysis (GSEA), tumor mutation burden (TMB), and clinic pathological relation analyses were also performed. RESULTS: Stromal, immune, and microenvironment comprehensive features (ESTIMATE score) were associated with overall survival (OS) in LUAD patients (all, P<0.05). T-cell activation, chemokine activity, and immune effect or dysfunction gene ontology maps were associated with the LUAD immune microenvironment. The immune infiltration cell subtypes mast cells (masT-cells) resting [The Cancer Genome Atlas (TCGA): P=0.01; Gene Expression Omnibus (GEO): P=1.79e-05] and activated T-cells (CD4 memory) (TCGA: P<0.01; GEO: P=8.52e-05) were found to have an important role in the immune cell regulatory network. Finally, ITGAL [univariate hazard ratio (HR) =0.80, 95% confidence interval (CI): 0.69-0.93, P<0.01; multivariate HR =0.59, 95% CI: 0.40-0.86, P=0.01] and KLRB1 (univariate HR =0.78, 95% CI: 0.69-0.89, P<0.01; multivariate HR =0.72, 95% CI: 0.58-0.90, P<0.01) were correlated with the T-cell receptor signaling pathway and anaplastic lymphoma kinase (ALK) fusion (ITGAL: P=0.034; KLRB1: P=0.050), and were considered as candidate biomarkers. A significant relation between KLRB1 expression level and TMB (P=3.6e-05) was identified, while no relation was detected for ITGAL (P=0.11). CONCLUSIONS: The T-cell activation and activated T-cell (CD4 memory) pathways were predominantly involved in LUAD immune microenvironment regulation. The expression levels of ITGAL and KLRB1 were significantly correlated with the T-cell receptor signaling pathway and LUAD TMB, and were independent risk factors for OS.

Nomograms Predict Overall Survival and Cancer-Specific Survival in Patients with Fibrosarcoma: A SEER-Based Study.

PURPOSE: Due to the rarity, it is difficult to predict the survival of patients with fibrosarcoma. This study aimed to apply a nomogram to predict survival outcomes in patients with fibrosarcoma. METHODS: A total of 2235 patients with diagnoses of fibrosarcoma were registered in the Surveillance, Epidemiology, and End Results database, of whom 663 patients were eventually enrolled. Univariate and multivariate Cox analyses were used to identify independent prognostic factors. Nomograms were constructed to predict 3-year and 5-year overall survival and cancer-specific survival of patients with fibrosarcoma. RESULTS: In univariate and multivariate analyses of OS, age, sex, race, tumor stage, pathologic grade, use of surgery, and tumor size were identified as independent prognostic factors. Age, sex, tumor stage, pathologic grade, use of surgery, and tumor size were significantly associated with CSS. These characteristics were further included to establish the nomogram for predicting 3-year and 5-year OS and CSS. For the internal validation of the nomogram predictions of OS and CSS, the C-indices were 0.784 and 0.801. CONCLUSION: We developed the nomograms that estimated 3-year and 5-year OS and CSS. These nomograms not only have good discrimination performance and calibration but also provide patients with better clinical benefits.

A Nomogram for Prediction of Postoperative Pneumonia Risk in Elderly Hip Fracture Patients.

PURPOSE: Pneumonia is one of the common complications of hip fracture. This study aimed to evaluate the risk factors and apply a nomogram to predict postoperative pneumonia in elderly hip fracture patients. MATERIALS AND METHODS: From August 2014 to October 2019, 1113 hip fracture patients who were older than 65 years and underwent surgical treatment in our hospital were subjects of this study. Univariate and multivariate Cox analyses were used to identify independent risk factors. A predictive nomogram model was built, and the discrimination and calibration were determined by receiver operating characteristic and calibration plot. RESULTS: A total of 166 patients developed pneumonia after operation (14.91%, pneumonia group) while the remaining 947 patients did not (85.09%, non-pneumonia group). According to the results, body mass index (OR, 0.76, 95% CI, 0.70 to 0.84, P<0.001), serum albumin (OR, 0.86, 95% CI, 0.79 to 0.93, P<0.001), c-reactive protein (OR, 1.01, 95% CI, 1.00 to 1.92, P=0.011), functional status (OR, 2.94, 95% CI, 1.69 to 5.10, P<0.001) and time to surgery (OR, 4.56, 95% CI, 2.64 to 7.88, P<0.001) were identified as independent risk factors of pneumonia. The area under the curve value for postoperative pneumonia risk was 0.905, and the P-value of the Hosmer-Lemeshow calibration test was 0.529. CONCLUSION: Our nomogram model can be used to predict the risk of pneumonia in elderly hip fractures after surgery and provide clinicians with guidance for better perioperative intervention to improve prognosis and reduce mortality.

PTEN inhibits non-small cell lung cancer cell growth by promoting G0/G1 arrest and cell apoptosis.

Non-small cell lung cancer (NSCLC) is a major type of human lung cancer and the primary cause of cancer-associated cases of mortality worldwide. Phosphatase and tensin homolog (PTEN) is a potent tumor suppressor gene in various human cancer types. The aim of the current study was to explore the role of PTEN and its associated regulatory mechanisms in NSCLC. Firstly, the expression of PTEN was detected using western blotting in a variety of NSCLC cell lines. The results revealed that compared with normal control cells, PTEN levels were significantly decreased in NSCLC cell lines (P<0.01). Short hairpin (sh)RNAs specific to PTEN were also used to knockdown endogenous PTEN in NSCLC cells. The results indicated that cell viability was significantly increased in PTEN-knockdown cells compared with those transfected with negative control shRNA (P<0.01). Conversely, overexpression of PTEN in A549 and SK-MES-1 cells significantly decreased the optical density of NSCLC cells (P<0.01). Flow cytometry was used to investigate the cell cycle; the results revealed that PTEN knockdown significantly increased the percentage of cells at G0/G1 phase (P<0.01) and decreased the number of cells at S phase (P<0.01). The molecular mechanism was further explored using western blotting and the results demonstrated that PTEN overexpression increased the levels of cleaved caspase-3 (P<0.01). These results suggest that PTEN may be a potential target gene for gene therapy in patients with NSCLCs.

In Vitro Growth of Mouse Preantral Follicles Under Simulated Microgravity.

14 day-old mouse ovarian tissue and preantral follicles isolated from same-aged mice were incubated in a simulated microgravity culture system. We quantitatively assessed follicle survival, measured follicle and oocyte diameters, and examined ultrastructure of the oocytes produced from the system. We observed decreased follicle survival, downregulation of expressions of proliferating cell nuclear antigen and growth differentiation factor 9, as indicators for the development of granulosa cells and oocytes, respectively, and oocyte ultrastructural abnormalities under the simulated microgravity condition. The simulated microgravity experimental setup needs to be optimized to provide a model for investigation of the mechanisms involved in the oocyte/follicle in vitro development.

Specific immunotherapy generates CD8(+) CD196(+) T cells to suppress lung cancer growth in mice.

That specific immunotherapy can inhibit cancer growth has been recognized; its efficiency is to be improved. This study aimed to inhibit lung cancer (LC) growth in a mouse model by using an LC-specific vaccination. In this study, a LC mouse model was created by adoptive transplantation with LC cells. The tumor-bearing mice were vaccinated with LC cell extracts plus adjuvant TNBS or adoptive transplantation with specific CD8(+) CD196(+) T cells. The results showed that the vaccination with LC extracts (LCE)/TNBS markedly inhibited the LC growth and induced CD8(+) CD196(+) T cells in LC tissue and the spleen. These CD8(+) CD196(+) T cells proliferated and produce high levels of perforin upon exposure to LCE and specifically induced LC cell apoptosis. Exposure to TNBS induced RAW264.7 cells to produce macrophage inflammatory protein-3α; the latter activated signal transducer and activator of transcription 3 and further induced perforin expression in the CD8(+) CD196(+) T cells. Adoptive transfer with specific CD8(+) CD196(+) T cells suppressed LC growth in mice. In conclusion, immunization with LC extracts and TNBS can induce LC-specific CD8(+) CD196(+) T cells in LC-bearing mice and inhibit LC growth.

Intrathoracic extramedullary hematopoiesis presenting as tumor-simulating lesions of the mediastinum in α-thalassemia: A case report.

Extramedullary hematopoiesis (EMH) is a rare disease, where hematological disorder drives extramedullary hematopoietic tumor formation in multiple regions of the body. The present study reports a case of EMH presenting as multiple tumor-like lesions of mediastinum in a 61-year-old male with α-thalassemia, which was subjected to a video-assisted thoracoscopic surgery tissue biopsy to differentiate it from other mediastinal tumors. To date, only three cases of EMH in patients with α-thalassemia have been described in the literature. Patients with EMH typically exhibit no hematological disorder preoperatively and therefore EMH is frequently misdiagnosed. In the present study, along with a literature review of the clinicopathological features of EMH, the diagnosis and treatment of this rare case was discussed, in order to differentiate diagnosis, and particularly to distinguish EHM from extramedullary myeloid sarcoma.

Low miR-145 silenced by DNA methylation promotes NSCLC cell proliferation, migration and invasion by targeting mucin 1.

MiR-145 has been implicated in the progression of non-small cell lung cancer (NSCLC); however, its exact mechanism is not well established. Here, we report that miR-145 expression is decreased in NSCLC cell lines and tumor tissues and that this low level of expression is associated with DNA methylation. MiR-145 methylation in NSCLC was correlated with a more aggressive tumor phenotype and was associated with poor survival time, as shown by Kaplan-Meier analysis. Additional multivariate Cox regression analysis indicated that miR-145 methylation was an independent prognostic factor for poor survival in patients with NSCLC. Furthermore, we found that restoration of miR-145 expression inhibited proliferation, migration and invasion of NSCLC by the direct targeting of mucin 1 by miR-145. Our results indicate that low miR-145 expression, due to methylation, promotes NSCLC cell proliferation, migration and invasion by targeting mucin 1. Therefore, miR-145 may be a valuable therapeutic target for NSCLC.

Overexpression of P21-activated kinase 4 is associated with poor prognosis in non-small cell lung cancer and promotes migration and invasion.

BACKGROUND: P21-activated kinase 4 (PAK4), an effector of the Rho family protein Cdc42, is an important oncogene whose expression is increased in many human cancers and is generally positively correlated with advanced disease and decreased survival. However, little is known about the expression and biological function of PAK4 in human non-small cell lung cancer (NSCLC). METHODS: PAK4 expression in NSCLC tissues and adjacent non-tumor tissues were assessed by immunohistochemistry, real-time PCR, and western blotting. Prognostic value of PAK4 expression was evaluated by Kaplan-Meier analysis and Cox regression. siRNA-mediated gene silencing and protein kinase assay was applied to demonstrate the role and the mechanism of PAK4 in lung cancer cell migration, invasion. RESULTS: The results showed that PAK4 was overexpressed in NSCLC cell lines and human NSCLC tissues. PAK4 expression was detected both in the membranes and cytoplasm of NSCLC cancer cells in vivo. Moreover, increased expression of PAK4 was associated with metastasis, shorter overall survival, advanced stage of NSCLC. Furthermore, PAK4 expression was positively correlated with phosphorylation of LIMK1 expression levels. Knockdown of PAK4 in NSCLC cell lines led to reduce the phosphorylation of LIMK1, which resulted in decrease of the cell migration and invasion. In addition, PAK4 bound to LIMK1 directly and activated it via phosphorylation. CONCLUSIONS: These data demonstrate that PAK4 mediated LIMK1 phosphorylation regulates the migration and invasion in NSCLC. Therefore, PAK4 might be a significant prognostic marker and potential therapeutic molecular target in NSCLC.

Multipotent Adult Progenitor Cells from Bone Marrow Differentiate into Chondrocyte-Like Cells.

Cartilage tissue engineering has great potential for treating chondral and osteochondral injuries. Efficient seed cells are the key to cartilage tissue engineering. Multipotent adult progenitor cells (MAPCs) have greater differentiation ability than other bone-marrow stem cells, and thus may be candidate seed cells. We attempted to differentiate MAPCs into chondrocyte-like cells to evaluate their suitability as seed cells for cartilage tissue engineering. Toluidine blue and Alcian blue staining suggested that glycosaminoglycan was expressed in differentiated cells. Immunofluorostaining indicated that differentiated human MAPCs (hMAPCs) expressed collagen II. Based on these results, we concluded that bone-marrow-derived hMAPCs could differentiate into chondrocyte-like cells in vitro.

Effect of lowering or restricting sympathectomy levels on compensatory sweating.

OBJECTIVE: There is controversy on whether lowering or restricting the level of sympathectomy can reduce compensatory sweating (CS). This study compared the results from sympathectomies performed to treat severe palmar hyperhidrosis using two distinct levels of T2-4 and T3-4. METHODS: One hundred and sixteen patients with primary palmar hyperhidrosis were randomly allocated to undergo either T2-4 sympathectomy treatment (T2-4 group) or T3-4 sympathectomy treatment (T3-4 group). Follow-up data were collected using a telephone questionnaire to assess efficacy, side effects, overall satisfaction, and factors affecting CS and the degree of satisfaction. RESULTS: There were no significant differences with respect to either CS or severe CS between the two treatment groups at 1, 6, or 12 months of follow-up. The total scores of the quality-of-life questionnaires after surgery were remarkably decreased compared with those before surgery in the two groups. However, no significant differences in quality-of-life scores were found between the two groups before surgery, or at 1, 6, or 12 months of follow-up. Age was predictive of severe CS at 6 months of follow-up (P = 0.045). Severe CS was inversely associated with patient satisfaction at 1, 6, and 12 months of follow-up. INTERPRETATION: The issue of whether lowering or restricting the level of sympathectomy reduces CS is controversial and needs more supportive evidence. Age may be a predictive factor for severe CS at 6 and 12 months of follow-up. Severe CS is the only known factor that affects patient satisfaction, and family history may also be associated with patient satisfaction.

PLCE1 suppresses p53 expression in esophageal cancer cells.

The apoptotic mechanism dysfunction plays a critical role in cancer cell growth and escaping from cancer therapies; the underlying mechanisms are to be further elucidated. This study aims to investigate the role of phospholipase C epsilon 1 (PLCE1) in modulating the apoptosis mechanism in esophageal cancer (Eca) cells. The results showed that Eca cell lines, OE33 and CP-C cells expressed high levels of PLCE1. Knockdown of PLCE1 markedly increased 9.26 folds of the expression of p53 and 13.8 folds of the frequency of apoptotic CP-C cells via modulating the p53 promoter methylation.

[Predicting efficacy of neoadjuvant cheomotherapy on resectable stage IIIA non-small cell lung cancer by multi-gene expressions].

BACKGROUND & OBJECTIVE: Neoadjuvant chemotherapy for stage IIIA non-small cell lung cancer (NSCLC) remains controversial. The role of the expressions of P53, K-ras, HER2, vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), CD44, and matrix metalloproteinase-9 (MMP-9) in predicting efficacy of neoadjuvant chemotherapy on stage IIIA NSCLC is still unclear although they have been found to be related to prognosis. This study was to determine the predictive effect of multi-gene expression on treatment outcome of neoadjuvant chemotherapy for resectable stage IIIA NSCLC. METHODS: Expressions of p53, K-ras, HER2, VEGF, EGFR, CD44, MMP-9 in the patients enrolled in the prospective randomized controlled trial were detected by immunohistochemistry. The treatment efficacies of combination (neoadjuvant chemotherapy combined with surgery) group (36 patients) and surgery alone group (32 patients) were compared. RESULTS: The high gene expression rate was 58.3% in combination group, and 40.6% in surgery alone group(P=0.145). In combination group, no significant difference of disease-free survival rate (P=0.903) and survival time (P=0.238) was found between patients with histopathologic regression and patients without histopathologic regression; high gene expression had no correlation with pathologic regression (P= 0.862); the mean disease-free survival time was significantly lower in high gene expression subgroup than in low gene expression subgroup [(14.1+/-9.8) months vs. (27.2+/-13.6) months, P=0.032]; the 2-year disease-free survival rate was 38.1% in high gene expression subgroup, and 46.7% in low gene expression subgroup (P=0.607). CONCLUSIONS: Pathologic regression after neoadjuvant chemotherapy has no correlation with disease-free survival rate and survial time. The high gene expression maybe indicate high risk of postoperative metastasis; the necessity of postoperative chemotherapy needs further study.

[Treatment of non-small-cell lung cancer with paraplatin given by two different dosage calculation methods].

OBJECTIVE: To study the reasonable dosage for paraplatin according to different dosage calculations. METHODS: A prospective, randomized, single-blinded study on 54 patients with advanced non-small-cell lung cancer (NSCLC) treated with paraplatin was conducted. Patients were divided to 2 groups. In group A, paraplatin dosage was calculated according to patients' body surface, and in group B, it was calculated according to the area under the curve (AUS). Hematological toxicity, response rate and survival rate in the two groups of patients were compared. RESULTS: Neutropenia in group A and group B was seen in 77.8% and 37.0% (P < 0.05), and thrombocytopenia in 18.5% and 3.7% (P > 0.05) of patients, respectively. Hemoglobin decrease was seen in 48.2% of patients in both groups. The average quantity of paraplatin given in one cycle of treatment was 535.93 +/- 106.71 mg and 398.52 +/- 71.72 mg (P < 0.01) respectively. The average time interval between treatment cycles was 27.04 +/- 5.30 d and 22.85 +/- 2.80 d (P < 0.05). The response rate and survival rate of patients in group A and B were 22.2% versus 48.2% (P < 0.05), and 40.7% versus 44.4% (P > 0.05) respectively, but the median survival time was identical (12 months) in the two groups. CONCLUSION: NSCLC patients given paraplatin with dosages calculated on the basis of AUC have higher response rate and less severe hematological toxicity than those given paraplatin with dosages on the basis of body surface. However, the median survival time and survival rate have no statistical differences between the two groups of patients.

[The application of needle video-assisted thoracoscopic biopsy in pathologic diagnosis and staging for advanced lung cancer].

OBJECTIVE: To study the clinical application of needle video-assisted thoracoscopic biopsy in the pathologic diagnosis and staging for advanced lung cancer. METHODS: Ninety-four patients were diagnosed as having advanced lung cancer staged IIIa-IV by chest X-ray, computed tomography and magnetic resonance imaging, for which a pathologic diagnosis was not made by sputum and pleural effusion cytology or bronchoscopic examination. Needle video-assisted thoracoscopic biopsy was performed for primary lesions, mediastinal lymph nodes, metastatic lesions in the lungs and the chest wall. RESULTS: Pathologic samples were obtained in 89 of the 94 patients by needle video-assisted thoracoscopic surgery. The successful rate was 95%. After operation, the pathologic diagnosis was confirmed to be lung cancer in all the 89 patients. Adenocarcinoma was found in 47 patients, squamous carcinoma in 23, adenosquamous carcinoma in 12, and small cell lung cancer in 7. Compared to the clinical diagnosis before operation, the pathologic diagnosis post-operation was changed in 15 patients. Pneumothorax and mild haemoptysis occurred in 4 patients and 2 of the cases respectively. All the patients were followed for 8 - 18 months with computed tomography and physical examination. No implantation metastasis was found. CONCLUSION: Needle video-assisted thoracoscopic biopsy is an effective diagnostic measure for patients with advanced lung cancer for which pathologic diagnosis, typing and staging are not determined by routine examinations.

[The preliminary results of a phase II randomized clinical trial of high-dose toremifene chemosensitization in stage IIIB/IV non-small cell lung cancer].

BACKGROUND: To investigate whether high-dose toremifene can enhance the efficacy of chemotherapy in non small cell lung cancer. METHODS: Untreated stage IIIB/IV non-small cell lung cancer patients were randomly devided into group A (high-dose toremifene combined with the platinum-based chemotherapy) or group B (the same platinum-based chemotherapy alone). RESULTS: A total of 30 eligible patients had been recruited. Hemotologic and nonhemotologic toxicities were similar with no statistic difference. The median survival for group A was 8 months, 95% CI (6.63-9.37) versus 7.5 months, 95% CI (4.75-10.25) for group B ( P =0.9). One year-survival rate was 31% for group A versus 28% for group B ( P =0.87). The response rate was 25% for group A versus 21% for group B ( P =0.99). CONCLUSIONS: The results suggest that high-dose toremifene does not enhance the efficacy of platinum-based chemotherapy for IIIB/IV non-small cell lung cancer but toxicities are well tolerated.