Elevation of the Chemokine Pair CXCL10/CXCR3 Initiates Sequential Glial Activation and Crosstalk During the Development of Bimodal Inflammatory Pain after Spinal Cord Ischemia Reperfusion.

BACKGROUND/AIMS: Spinal microglia and astrocytes are the main responders to the inflammatory cascade and process pain through various neural interactions. CXCL10 is a late-phase protein that accelerates arteriogenesis during reperfusion through CXCR3. However, the early-phase expression (within 72 h postoperatively) of CXCL10 and CXCR3 during the development of ischemia-reperfusion (IR)-induced inflammatory pain remains unclear. We investigated whether this chemokine pair participates in glial interactions during early-phase IR injury. METHODS: A rat model was induced by an 8-min occlusion of the aortic arch. Temporal assessments of mechanical and thermal allodynia and the protein levels of CXCL10 and CXCR3 were determined through measurements of paw withdrawal thresholds (PWTs) and paw withdrawal latencies (PWLs) and Western blotting assays. The co-localization of various cells with glial cells was detected by double immunofluorescence. The effects of CXCL10/CXCR3 on glial interactions were explored by intrathecal treatment with specific inhibitors (AMD487, minocycline and fluorocitrate) and recombinant CXCL10, and subsequent release of cytokines was assessed by ELISAs. RESULTS: The IR injury initiated bimodal allodynia within 72 h of reperfusion, as illustrated by two W-shape trends in the PWTs and PWLs with two minima at 12 and 48 h post-IR. Allodynia was highly correlated with overexpression of CXCL10 and CXCR3, which were expressed in microglia at the early stage and in both microglia and astrocytes at the late stage, as shown by increased CXCL10 and CXCR3 immunoreactivities and double-labeled cells. AMD487 and minocycline injections exerted comparable inhibitory effects on CXCR3 and Iba-1 and on GFAP immunoreactivity at 12 and 48 h post-IR, and these inhibitory effects were only observed at 48 h following fluorocitrate injection. The levels of TNF-α and IL-6 showed variations in concert with the changes in Iba-1 and GFAP immunoreactivities. Recombinant CXCL10 injection reversed the abovementioned effects. CONCLUSION: The results showed that CXCL10/CXCR3 are involved in bimodal inflammatory pain during early-phase IR injury. The sequential activation of and crosstalk between microglia and astrocytes mediated through CXCR3 upregulation suggested that treatments targeting specific cell types are important in post-IR allodynia.

Serum Fork-Head Box D3 (FOXD3) Expression Is Down-Regulated in and Associated with Diagnosis of Patients with Non-Small Cell Lung Cancer.

BACKGROUND The aim of this study was to detect the expression of fork-head box D3 (FOXD3) and investigate its diagnostic value in patients with non-small cell lung cancer (NSCLC). MATERIAL AND METHODS The relative expression of FOXD3 at mRNA and protein levels was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting analysis, respectively. Chi-square test was used to explore the relevance of FOXD3 expression with clinical features of NSCLC patients. A receiver operating characteristic (ROC) curve was built to estimate the diagnostic value of FOXD3 in distinguishing NSCLC patients from healthy controls. RESULTS Serum FOXD3 expression was weakly expressed in NSCLC patients compared to the controls at mRNA and protein levels (P<0.001) and low FOXD3 expression was positively correlated with TNM stage, lymph node metastasis, and differentiation. The ROC curve indicated that FOXD3 acts as a diagnostic bio-marker for NSCLC patients, with an AUC of 0.826 corresponding to a sensitivity of 77.1% and a specificity of 74.6%, and an optimal cutoff point of 2.38. CONCLUSIONS Decreased expression of serum FOXD3 was observed in NSCLC patients, and it was found to be a potential molecular marker for the diagnosis of NSCLC.

Deficiency of Functional Iron-Sulfur Domains in ABCE1 Inhibits the Proliferation and Migration of Lung Adenocarcinomas By Regulating the Biogenesis of Beta-Actin In Vitro.

BACKGROUND/AIMS: ATP-binding cassette transporter E1 (ABCE1), a unique ABC superfamily member that bears two Fe-S clusters, is essential for metastatic progression in lung cancer. Fe-S clusters within ABCE1 are crucial for ribosome dissociation and translation reinitiation; however, whether these clusters promote tumor proliferation and migration is unclear. METHODS: The interaction between ABCE1 and ß-actin was confirmed using GST pull-down. The lung adenocarcinoma (LUAD) cell line A549 was transduced with lentiviral packaging vectors overexpressing either wild-type ABCE1 or ABCE1 with Fe-S cluster deletions (ΔABCE1). The role of Fe-S clusters in the viability and migration of cancer cells was evaluated using clonogenic, MTT, Transwell and wound healing assays. Cytoskeletal rearrangement was determined using immunofluorescent techniques. RESULTS: Fe-S clusters were the key domains in ABCE1 involved in binding to ß-actin. The proliferative and migratory capacity increased in cells overexpressing ABCE1. However, the absence of Fe-S clusters reversed these effects. A549 cells overexpressing ABCE1 exhibited irregular morphology and increased levels of cytoskeletal polymerization as indicated by the immunofluorescence images. In contrast, cells expressing the Fe-S cluster deletion mutant presented opposing effects. CONCLUSION: These results demonstrate the indispensable role of Fe-S clusters when ABCE1 participates in the proliferation and migration of LUADs by interacting with ß-actin. The Fe-S clusters of ABCE1 may be potential targets for the prevention of lung cancer metastasis.

Expression of ATP binding cassette E1 enhances viability and invasiveness of lung adenocarcinoma cells in vitro.

ATP binding cassette E1 (ABCE1), a member of the family of ATP binding cassette transporters, has initially emerged as an RNase L inhibitor. As a highly conserved protein, it is involved in capsid assembly and translation processes of the human immunodeficiency virus as well as in tumor development and progression. Studies have shown that ABCE1 protein was overexpressed in lung carcinoma tissues and metastatic lymph nodes compared to normal lung tissues. However, little is known about the roles of ABCE1 in lung cancer. The present study investigated the biological effects of vector-mediated ABCE1 overexpression in lung cancer cells in vitro and examined the underlying molecular mechanisms. Overexpression of ABCE1 in the LTEP­a-2 lung adenocarcinoma cell line was achieved by transfection with a plasmid containing full­length ABCE1 cDNA. The ectopic expression of ABCE1 was shown to promote the viability and invasive capacity of lung cancer cells, and to in reduce p27 expression. However, overexpression of ABCE1 did not significantly affect the cell cycle distribution. In conclusion, the present study suggested that ABCE1 promotes the growth, invasion and metastasis of lung adenocarcinoma cells and may represent a potential biomarker and therapeutic target for lung cancer.

ABCE1 plays an essential role in lung cancer progression and metastasis.

ATP-binding cassette E1 (ABCE1) is a member of the ATP-binding cassette transporters and regulates a broad range of biological functions including viral infection, cell proliferation, and anti-apoptosis. We have previously shown that ABCE1 is a prognostic indicator for lung cancer, although the underlying mechanisms remain unclear. To investigate whether the ABCE1 gene contributes to the malignancy of lung tumors, we introduced ABCE1 into LTEP-a-2 lung adenocarcinoma cells. Ectopic ABCE1 expression promoted clonogenicity and anchorage-independent growth of LTEP-a-2 cells, while in a mouse xenograft tumor model, it had an augmentative effect on tumor growth and metastasis and reduced the expression of the tumor-suppressor gene growth arrest and DNA damage-inducible 45α (GADD45α). Moreover, apoptosis was not significantly influenced by ABCE1 in vitro. In summary, we have provided evidence that ABCE1 plays an essential role in the progression and metastasis of lung cancers and may represent a valuable therapeutic target for the management of lung tumor.

[Research and practice about risk control of shenfu injection].

Shenfu injection is developed by improving dosage form of ancient prescription "Shenfu Tang" and is mainly derived from extracts of both traditional Chinese medicine red ginseng and prepared lateral root of monkshood with polysorbate 80 as auxiliary material. Shenfu injection may be administered through intramuscular injection, intravenous drip or intravenous injection. It produces good effects in restoring Yang and rescuing patients from collapse, tonifying Qi and preventing exhaustion. It is mainly used to treat not only syncope and prostration resulting from sudden Yang collapse (infectious, hemorrhagic and water depletion shock etc), but also pavor, palpitation, dyspnea with cough, stomachache, diarrhea and arthralgia etc caused by deficiency of Yang (deficiency of vital energy). Research group has audited the monitored hospitals and has carried out postmarketing study of Shenfu solution from many aspects including literature review, spontaneous reporting system (SRS) and hospital information system (HIS) data analysis etc. A summary is shown below.

Overexpression of RNF146 in non-small cell lung cancer enhances proliferation and invasion of tumors through the Wnt/ß-catenin signaling pathway.

Studies have suggested a possible correlation between the newly identified E3 ubiquitin ligase ring finger protein 146 (RNF146) and tumor development. However, until now, studies on RNF146 have been restricted to poly(ADP-ribosyl)ation and ubiquitin ligation, whereas the role of RNF146 in tumor biology has rarely been reported. In the present study, the role of RNF146 in non-small cell lung cancer (NSCLC) was investigated. The results showed that the expression of RNF146 was increased in clinical lung cancer samples and cell lines. RNF146 expression correlated with tumor size, differentiation level, lymphatic metastasis, pTNM staging, and prognosis of patients in stage I. RNF146 expression was negatively correlated with Axin expression but positively correlated with the nuclear expression of ß-catenin in NSCLC tissues. RNF146 downregulated the expression of Axin in lung cancer cell lines and induced the expression and nuclear distribution of ß-catenin. Overexpression of RNF146 in NSCLC cell lines increased the levels of cyclinD1, cyclinE, and CDK4, promoted cell cycle G0/G1-S transitions, and regulated cell proliferation. Overexpression of RNF146 led to upregulated levels of matrix metalloproteinases 2 and 7 and enhanced lung cancer cell invasiveness, events that were mediated by the classical Wnt/ß-catenin signaling pathway. In summary, the data in the present study indicate that RNF146 regulated the development and progression of NSCLC by enhancing cell growth, invasion, and survival, suggesting that RNF146 may be a potential treatment target in NSCLC.

PRKACB is downregulated in non-small cell lung cancer and exogenous PRKACB inhibits proliferation and invasion of LTEP-A2 cells.

Protein kinase cAMP-dependent catalytic ß (PRKACB) is a member of the Ser/Thr protein kinase family and a key effector of the cAMP/PKA-induced signal transduction involved in numerous cellular process, including cell proliferation, apoptosis, gene transcription, metabolism and differentiation. In the present study, the expression pattern of PRKACB in non-small cell lung cancer (NSCLC) and the effect of PRKACB upregulation on cell proliferation, apoptosis and invasion were investigated. PRKACB mRNA and protein expression was analyzed in the NSCLC tissue and corresponding normal tissues of 30 cases, using quantitative RT-PCR and western blot analysis. A plasmid containing full-length PRKACB was transfected into LTEP-A2 cells to further investigate the effects of PRKACB overexpression on proliferation, apoptosis and invasion of the transfected cells, which were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, flow cytometry and Transwell assays. The results revealed that the NSCLC tissues exhibited much lower levels of PRKACB mRNA and protein compared with their corresponding normal tissues. The upregulation of PRKACB decreased the numbers of proliferative, colony and invasive cells, while the apoptotic rates of transfected cells were increased. These data indicate that PRKACB is downregulated in NSCLC tissues and that upregulation of PRKACB may be an effective way to prevent the progression of NSCLC.

Expression of p114RhoGEF predicts lymph node metastasis and poor survival of squamous-cell lung carcinoma patients.

The Rho-specific guanine nucleotide exchanging factor p114RhoGEF is involved in RhoA activation and cell motility. Previous studies suggest that altered expression of p114RhoGEF could contribute to cancer progression. We investigated an association of p114RhoGEF expression with progression and prognosis of non-small cell lung cancer (NSCLC). Immunohistochemistry was performed to detect p114RhoGEF expression in 105 NSCLC (34 adenocarcinoma and 71 squamous-cell carcinoma) and 32 normal lung tissues. We found that p114RhoGEF expression was upregulated in squamous-cell lung carcinoma and that p114RhoGEF expression was significantly higher in squamous-cell carcinoma than in adenocarcinoma or normal tissues (P<0.05, both). Expression of p114RhoGEF protein was significantly associated with lymph node metastasis of lung cancer (P<0.05), but not with patients' age, gender, tumor size, differentiation, or stage. Expression of p114RhoGEF protein was also associated with poor overall and event-free survival of squamous-cell lung carcinoma patients (P<0.05). Taken together, p114RhoGEF expression may be useful in predicting progression and survival of squamous-cell lung carcinoma patients. A future study will investigate whether p114RhoGEF can serve as a novel therapeutic target in squamous-cell lung cancer.

The biological regulation of ABCE1.

ATP binding cassette E1 (ABCE1) protein, also named RLI or HP68, is one member of ATP binding cassette superfamily. By forming a heterodimer with RNase L, ABCE1 protein inhibits the IFN-depended 2-5A/RNase L system and regulates a broad range of biological functions including viral infection, tumor cell proliferation, and antiapoptosis. As a highly conserved protein, recent studies have mainly focused that ABCE1 protein is involving in the fields of HIV virus capsids assembly and important roles in translation. Our manuscript will review the studies which revealed the biological regulation of ABCE1.

[Risk control system construction of Shenfu injection].

OBJECTIVE: To ensure the stable quality of Shenfu injection, control potential risk and reduce risk damage. METHOD: To screen and evaluate the risk in the production, GAP, research and development, circulation, clinical application, intellectual property, emergency treatment,control and prevente the potential risks. RESULT: The risk-control system of Shenfu injection has been constructed initially. CONCLUSION: It has a great significance for the establishment of traditional Chinese medicive injections' risk-control system. The keys of the risk-control in future are to make strategy, perfect the organization structure and collaborating among various departments.

Surgery for young patients with lung cancer.

OBJECTIVES: To investigate the relationship between the clinical features and prognosis in young patients with lung cancer who underwent resection. METHODS: Statistical analysis was employed on sex, age, symptoms, diagnosis, treatment and prognosis, in 92 young cases younger than 40 years old among 930 cases with primary lung cancer who underwent surgery from January 1978 to December 1996. RESULTS: There were 92 young patients with lung cancer, accounting for 9.89% of the total cases. They were 71 male and 21 female patients, with the ratio of 3.38:1. The histological types were 34 squamous cell carcinomas (37%), 30 adenocarcinomas (33%), 26 small cell carcinomas (28%), and two large cell carcinomas (2%). On TNM staging, there were 30 cases in stage I (32.6%), 30 in stage II (32.6%) and 32 in stage III (34.8%). Lobectomy was conducted in 54 patients (59%), pneumonectomy in 36 (39%) and wedge-shaped resection in two cases (2%). The rate of pneumonectomy in young patients was significantly higher than that of 18% in older patients (>40) with lung cancer (P<0.01). 57 patients (62%) received absolutely curative resection; 28 cases (30%), relatively curative resection; seven cases (8%), non-curative resection. The postoperative 5-year-survival was 46% (42/92), in comparison with 34% (288/838) in patients older than 40 receiving operation during the same period, with significant difference between the two groups (P<0.05). The 5-year-survivals in patients with squamous cell carcinoma, adenocarcinoma, small cell carcinoma and large cell carcinoma were 68% (23/34), 30% (9/30), 38% (10/26) and 0 (0/2), respectively. The survival in squamous cell cancer was markedly higher than in adenocarcinoma (P<0.01) and in small cell carcinoma (P<0.05). There was no significant difference between adenocarcinoma and small cell cancer. The 5-year-survivals in stage I, II and III were 63% (19/30), 53% (16/30) and 22% (7/32), respectively. There was no significant difference between stage I and II, while remarkable difference was found between stage I and III (P<0.01), and between stage II and III (P<0.05). The 5-year-survival in patients undergoing absolutely curative resection was 67% (38/57), and 14% (4/28) in patients with relatively curative resection, with significant difference (P<0.01). No patient survived longer than the 5th postoperative year in seven cases receiving non-curative resection. CONCLUSIONS: Young patients with lung cancer were more often seen in male than in female. Squamous cell carcinoma accounted for the most part, no statistical difference, however, compared with adenocarcinoma and small cell carcinoma. The pneumonectomy rate in young patients was remarkably higher than that in patients older than 40. The postoperative 5-year-survival in young patients was considerably higher than in patients older than 40 who underwent surgery during the same period. Favorable prognosis was seen in patients with squamous cell carcinoma and undergoing absolutely curative resection, while worse outcome in stage III cancer.