Vitamin D-mediated tsRNA-07804 triggers mitochondrial dysfunction and suppresses non-small cell lung cancer progression by targeting CRKL.

OBJECTIVE: tRNA-derived small RNAs (tsRNAs) are novel non-coding RNAs with various functions in multiple cancers. Nevertheless, whether vitamin D executes its function in mitochondrial dysfunction and non-small cell lung cancer (NSCLC) progression through tsRNAs remains obscure. METHODS: Differentially expressed tsRNAs between control and vitamin D-treated H1299 cells were acquired by small RNA sequencing. Cell and animal experiments were implemented to elucidate the impacts of vitamin D and tsRNA on mitochondrial dysfunction and NSCLC progression. Dual-luciferase reporter assay, quantitative real-time PCR, western blot and recovery experiments were applied to determine the mechanism of tsRNA in NSCLC. RESULTS: We discovered that vitamin D receptor resulted in decreased mitochondrial-related functions and vitamin D caused mitochondrial dysfunction of NSCLC cells. tsRNA-07804 was remarkably upregulated in vitamin D-treated H1299 cells. Functional experiments indicated that vitamin D led to mitochondrial dysfunction, repressed the proliferation, migration, invasion, and promoted apoptosis of H1299 cells via regulating tsRNA-07804. Mechanistically, tsRNA-07804 induced mitochondrial dysfunction and inhibited the malignancy of H1299 cells by suppressing CRKL expression. In vivo experiments showed that vitamin D inhibited the tumor growth in NSCLC by increasing tsRNA-07804 expression. Moreover, clinical sample analysis unveiled that tsRNA-07804 had a negative correlation with CRKL. CONCLUSIONS: In conclusion, our study proved that vitamin D induced mitochondrial dysfunction and suppressed the progression of NSCLC through the tsRNA-07804/CRKL axis. Overall, these results unveiled that tsRNA-07804 might act as a potential therapeutic target for NSCLC.

MiR-101-3p inhibits the growth and metastasis of non-small cell lung cancer through blocking PI3K/AKT signal pathway by targeting MALAT-1.

BACKGROUND: MiR-101-3p is an important tumor suppressor miRNA in many human cancers. This study was to investigate the role of miR-101-3p in the progression of non-small cell lung cancer (NSCLC) and its potential underlying mechanism. METHODS: In this study, the endogenous expression of miR-101-3p or MALAT-1 in cells was modulated by cell transfection assays. The regulatory interaction of miR-101-3p and MALAT-1 was examined by Luciferase reporter gene and RNA pull-down assays. The effect of miR-101-3p or MALAT-1 on NSCLC cells was evaluated by cell proliferation assays, wound-healing assays and transwell invasion assays. The mice tumor model was established to test the role of miR-101-3p and MALAT-1 in the growth and metastasis of NSCLC in vivo. RESULTS: The relative expression of miR-101-3p in NSCLC cells was significantly decreased; while MALAT-1 was significantly increased. Moreover, overexpression of miR-101-3p could significantly inhibit the proliferation, migration and invasion of NSCLC cells in vitro, as well as MALAT-1 expression. Further studies confirmed that miR-101-3p could specifically repress MALAT-1 expression through direct binding; MALAT-1 overexpression completely reversed the miR-101-3p-induced suppression on the viability, migration and invasion of NSCLC cells and MALAT-1 expression. Finally, we confirmed that miR-101-3p could block the MALAT-1-induced activation of PI3K/AKT signal pathway and resulted in the inhibition on the growth and metastasis of NSCLC cells in vivo. CONCLUSION: MiR-101-3p inhibited the growth and metastasis of NSCLC through blocking PI3K/AKT signal pathway by targeting MALAT-1.

Efficacy and Safety of Belimumab Plus Standard Therapy in Patients With Systemic Lupus Erythematosus: A Meta-analysis.

PURPOSE: The treatment of belimumab plus standard therapy in patients with systemic lupus erythematosus (SLE) has been studied extensively in recent years. Our aim was to estimate the efficacy and safety of this therapy compared with placebo plus standard therapy in patients with SLE. METHODS: PubMed, Web of Science, Embase, Chinese Biomedical Literature Database (CBM, Chinese), and Wanfang Database (Chinese) were searched for all randomized clinical trials that mainly studied the efficacy and safety of belimumab plus standard therapy before June 2015. We extracted or calculated the rate of the SLE Response Index and adverse event rate at 52 weeks in all the included studies. The odds ratio (OR) with 95% CI between the 2 groups in this meta-analysis was conducted by using a random-effects model. Sensitivity and publication bias analyses were also performed. All statistical tests were performed by using Stata software version 12.0 (StataCorp., College Station, Texas). FINDINGS: In the overall samples (4 studies, N = 4692 ), a significantly higher SLE Response Index rate at 52 weeks was found in belimumab plus standard therapy group compared with the placebo plus standard therapy group in all studies (OR = 1.49; 95% CI, 1.26-1.77 ; P < 0.001 ). When assessed with the incidence of serious adverse events, the data revealed that there was no significant difference between the 2 groups, with pooled OR = 1.08; 95% CI, 0.83-1.39; P = 0.573; OR = 1.23; 95% CI, 1.02-1.48; P = 0.029; and OR = 1.07; 95% CI, 0.88-1.29; P = 0.506. IMPLICATIONS: The results suggest that treatment with belimumab plus standard therapy is more effective than placebo plus standard therapy in SLE patients, which represents major progress in the treatment of SLE. Regardless of the statistical analyses, further research is necessary to optimize treatment effects.

Prognostic value of matrix metalloproteinase-7 expression in patients with non-small cell lung cancer.

The prognostic value of matrix metalloproteinase-7 (MMP-7) for survival of patients with non-small cell lung cancer (NSCLC) remains controversial. We performed a meta-analysis of the literatures to clarify its impact. Trials were selected for meta-analysis if they provided an independent assessment of MMP-7 in NSCLC and reported the analysis of survival data based on MMP-7 status. Pooled hazard ratio (HR) with 95% confidence interval (95% CI) was used to evaluate the associations between MMP-7 expression and survival of NSCLC patients. Heterogeneity and publication bias were also assessed. Seven studies involving 1,446 patients were identified. The combined HR for all studies was 1.28 (95% CI 0.86-1.91; P = 0.22). Subgroup analysis revealed that MMP-7 overexpression had a favorable impact on survival in Caucasians (HR = 0.74; 95% CI 0.55-0.99; P = 0.043) but showed a poor survival prognosis in Asians (HR = 1.74; 95% CI 1.05-2.88, P = 0.031). Its effect also appeared significant when the analysis was restricted to Asian patients with squamous cell cancer (HR =3.42; 95% CI 1.92-6.11, P = 0.000) and adenocarcinoma (HR = 2.1; 95% CI 1.34-3.29, P = 0.001). Our meta-analysis suggests that there are ethnic differences in the clinical significance of MMP-7 expression for patients with NSCLC.

Surgical strategy for tubercular abscess in the chest wall: experience of 120 cases.

OBJECTIVES: We aim to optimize surgical strategy to decrease relapse of tubercular abscess in the chest wall (TACW). METHODS: The records of 120 patients who underwent surgical treatment for TACW from May 2005 to March 2011 were retrospectively reviewed. We conducted the following surgical treatment as '6C + A' by abbreviating the first alphabet of each step: (i) careful exploration of the abscess; (ii) complete resection; (iii) cavity washing using sodium bicarbonate solution; (iv) coverage using muscle flap; (v) continuous suction and drainage; (vi) compression dressing and (vii) anti-tuberculosis medication. RESULTS: One hundred and thirteen cases were discharged for rehabilitation with the first stage wound healing (113/120). Four cases postoperatively suffered from subcutaneous fistula which was healed after dressing changes for 1-2 months. Three patients with an abscess relapse underwent the second operation 2 months after the first operation. Follow-ups ranged from 2 months to 6 years and demonstrated no recurrence. CONCLUSIONS: We deem the surgical procedures '6C + A' effective to obviate relapse of TACW.

Clinical value of assaying tumor supplied group of factor/tumor specific growth factor in patients with solitary pulmonary nodule.

OBJECTIVE: This pilot study was designed to evaluate the clinical value of assaying tumor supplied group of factor/tumor specific growth factor (TSGF) in solitary pulmonary nodule (SPN). PATIENTS AND METHODS: The study was conducted from March 2007 to September 2010 and included 33 patients with SPN and 28 healthy volunteers. TSGF was assayed in preoperative serum, intraoperative pleural lavage fluid (IPLF), and postoperative serum. RESULTS: At operation, 20 patients were diagnosed with malignancy and 13 patients were diagnosed with nonmalignancy and placed in group A and group B, respectively. In group A, pathologic staging demonstrated 8 patients (group A1) with stage T1N0M0, 7 patients (group A2) with stage T1N1M0 and 53 patients (group A) with stage T1N2M0 disease. In group B, 8 patients were diagnosed with tuberculoma (group B1) and 5 patients were diagnosed with inflammatory pseudotumor (group B2). Before operation, levels of TSGF in peripheral blood were significantly higher in group A compared with group B and the control group (98.8 ± 29.9 vs. 62.1 ± 24.9 and 50.1 ± 17.9, Student-Newman-Keuls test; P < .05). The percentage of patients with positive serum TSGF results was significantly higher in group A than in group B or the control group (90.0% vs. 30.8% and 17.9%, χ(2) test; P < .05). With respect to the diagnostic value of serum TSGF in malignant SPN, we found sensitivity to be 90%, specificity to be 69.2%, positive forecast rate to be 74.5%, negative forecast rate to be 87.4%, and accurate diagnosed rate to be 79.5%. The TSGF level in IPLF in group A was significantly higher than that in group B (132.2 ± 51.9 vs. 84.6 ± 12.6, Student t test, P < .05). Additionally, TSGF in group A2 and group A3 was significantly higher compared with group A1 (162.2 ± 52.3 and 176.4 ± 17.8 vs. 100.2 ± 35.8, Student-Newman-Keuls test; P < .05). Postoperative serum TSGF in the patients diagnosed with lung cancer decreased significantly after operation. TSGF returned to a normal threshold level (71 U/mL) in the sixth month postoperatively. In addition, there was no appreciable change in the patients in group B. CONCLUSION: Serum TSGF is conducive to discriminating between benign and malignant features of SPN. Additionally, investigation of IPLF TSGF can potentially offer a new approach to predict the existence of lymph node metastases.

Breast cancer anti-estrogen resistance protein 1 (BCAR1/p130cas) in pulmonary disease tissue and serum.

OBJECTIVE: The purpose of the study was to evaluate clinical presentation of breast cancer anti-estrogen resistance protein 1 (BCAR1, also known as p130cas) expression in pulmonary diseases, and to assess its potential as a molecular marker for diagnosis and prognosis. METHODS: Between March 2008 and August 2010, we enrolled a total of 80 patients (group A) with non-small-cell lung cancer (NSCLC), 48 patients (group B) with pulmonary tuberculosis (including 27 cases of tuberculoma and 21 cases of cavitary pulmonary tuberculosis), and 32 patients (group C) with other benign pulmonary mass (hamartoma in 15 cases, inflammatory pseudotumor in 10 cases, fibroid tumor in 7 cases). Additionally, 160 healthy age- and sex-matched volunteers were recruited as healthy controls. Tissue BCAR1 expression was investigated by using tissue microarray and immunohistochemistry. BCAR1 and tumor markers (carcinoma embryonic antigen [CEA] and the cancer antigens CA19-9 and CA125) in serum were assayed by using ELISA and immunoradiometrics, respectively. RESULTS: BCAR1 expression was detected (either in the nucleus, the cytoplasm, or both) in tumor cells in 79 of the 80 NSCLC cases in group A, and in fibroblasts in 41 of the 48 pulmonary tuberculosis cases in group B. However, it was not detected in the normal adjacent tissue in 70 of the 80 cases in group A and in 47 of the 48 cases in group B. In group C, BCAR1 expression was negative in all 32 cases. Additionally, we investigated adjacent tissue with acute or chronic inflammation in 20 cases from group C, and found no expression of BCAR1. Serum BCAR1 levels were significantly higher in patients with NSCLC than in the control group, increased gradually with the progression of tumor staging, and decreased after removal of the tumors. The levels were significantly lower in bronchioloalveolar carcinoma than in other subtypes of carcinoma (Mann-Whitney U test, Z = -5.089; p < 0.001). Serum BCAR1 levels were significantly higher in patients with pulmonary tuberculosis than in the control group, were positively and significantly correlated with the diameter of the tuberculosis lesion (Spearman's rho, correlation coefficient 0.753; p < 0.001), and decreased after removal of the tuberculosis lesions. The levels were significantly higher in patients with cavitary pulmonary tuberculosis than in those with tuberculoma (517.6 ± 326.5 vs 282.2 ± 137.6; Student's t-test, t = -3.387; p = 0.001). In group C, there was no appreciable difference in serum BCAR1 levels compared with the matched controls (222.8 ± 111.0 vs 201.6 ± 35.7; Dunnett's T3 test, p = 0.993). The discrimination power of combining BCAR1 and tumor markers in NSCLC versus benign lung diseases was higher than that of sole use of BCAR1 as a marker (maximal sum of sensitivity and specificity: 1.538 vs 1.237). CONCLUSION: We conclude that a combined assay of serum BCAR1 and traditional tumor markers is potentially applicable for distinguishing NSCLC from benign lung diseases. However, the clinical utility of serum BCAR1 as a molecular marker for prognosis in NSCLC or pulmonary tuberculosis requires further clarification and verification.

The impact of acute atrial fibrillation on the prothrombotic state in patients with essential hypertension.

OBJECTIVES: To investigate whether acute atrial fibrillation (AF) creates a prothrombotic state in hypertensive patients, and to assess the evolution in research indices after cardioversion. DESIGN AND METHODS: Plasma levels of von Willebrand factor (vWf), soluble P-selectin and fibrin D-dimers were measured in hypertensive patients with acute AF pre-cardioversion and at 1, 7, 14, and 30days post-cardioversion. The results were compared with hypertensive controls and healthy controls. RESULTS: Pre-cardioversion plasma vWf levels in acute AF patients were similar to those of controls; however, post-cardioversion, the vWf levels increased significantly and only returned to baseline levels 14days later. Compared with controls, acute AF patients had higher levels of fibrin D-dimers, which lasted at least 14days after cardioversion. CONCLUSIONS: This study demonstrated that hypertensive patients with acute AF have an abnormal prothrombotic state, which persists for up to 14days after cardioversion.

Gene silencing of BAG-1 modulates apoptotic genes and sensitizes lung cancer cell lines to cisplatin-induced apoptosis.

BAG-1 is a multifunctional protein that interacts with a wide range of target molecules to regulate apoptosis. However, the exact mechanism by which BAG-1 inhibits apoptosis is unknown. In the current study, we comprehensively investigate BAG-1 mediated changes in gene expression by comparing the gene expression profile after BAG-1 siRNA silencing in the human lung adenocarcinoma cell line A549. Microarray assay revealed that expression of 589 genes was altered, including 177 upregulated and 412 downregulated genes. Real-time PCR assay of the changes in gene expression indicated a concordance of 86.48% (64/74) with that observed by microarray. The upregulated genes included EGFR, Jun, HSPA1A and CASP7; while the downregulated genes included Lyn, HRAS and TGFalpha. Furthermore, we confirmed by western blot that the expression of EGFR, Jun and HSPA1A were significantly increased, while expression of Lyn and HRAS were dramatically decreased after BAG-1 gene silencing. After transfection with BAG-1 siRNA, lung cancer cell lines A549 and L9981 exhibited significantly enhanced sensitivity to cisplatin-induced apoptosis. Thus, silencing of the BAG-1 gene resulted in changes in expression of apoptosis-related genes and sensitized A549 and L9981 cells to cisplatin-induced apoptosis.

MMP-2 and TIMP-2 gene polymorphisms and susceptibility to atrial fibrillation in Chinese Han patients with hypertensive heart disease.

BACKGROUND: MMP-2 and TIMP-2 play important roles in the pathogenesis of arrhythmogenic atrial remodeling, and may contribute to the development and persistence of atrial fibrillation (AF). Functional polymorphisms in the promoter of MMP-2 and TIMP-2 gene may modulate an individual's susceptibility to AF. METHODS: A total of 881 hypertensive heart disease patients from Chinese Han population (128 with and 753 without AF) were recruited in this study. The genotypes of the MMP2-1306C>T and -735C>T polymorphisms and TIMP-2 -418G>C polymorphisms were determined using PCR based method. The plasma concentration of TIMP-2 was measured by enzyme-linked immunosorbent assay in a subgroup with 81 patients. RESULTS: Both genotype distribution and allele frequency of the TIMP-2 -418G>C polymorphism were significantly different between the AF and control group (P=0.005 and P=0.001, respectively). The C allele carriers (GC+CC) had a significantly increased risk of AF compared with the GG homozygotes (odds ratio,1.77, 95% CI 1.21-2.92, P=0.009) in a logistic regression model after adjustment for age, left atrial dimension, left ventricular mass index, and antihypertensive drugs. The C allele carriers also had reduced levels of plasma TIMP-2 levels compared with GG homozygotes in both AF patients and control subjects. No relationship was found in this cohort between the presence of the MMP-2 -1306C>T and -735C>T polymorphism and AF. CONCLUSIONS: The TIMP-2 -418G>C polymorphism is significantly associated with an increased susceptibility to AF in Chinese Han patients with hypertensive heart disease. The -418C allele, which is associated with a decreased expression of TIMP-2, might be a genetic risk for the development of AF in this cohort.

Association of MMP-9 gene polymorphisms with atrial fibrillation in hypertensive heart disease patients.

BACKGROUND: MMP-9 plays an important role in the pathogenesis of arrhythmogenic atrial remodeling, and may contribute to the development and persistence of atrial fibrillation (AF). Functional polymorphisms in the MMP-9 gene which lead to altered MMP-9 production and/or activity may modulate an individual's susceptibility to AF. METHODS: A total of 881 hypertensive heart disease patients of Chinese Han population (128 with and 753 without AF) were recruited in this study. The MMP-9 -1562C>T and R279Q genotypes were determined using PCR-RFLP method. The plasma concentration of MMP-9 was measured by ELISA. RESULTS: Both the genotype distributions and allele frequencies of the -1562C>T polymorphism were significantly different between the AF and control group (P=0.007 and P=0.002, respectively). The T allele carriers (TT + CT) had significantly increased risk of AF compared with the CC homozygotes (OR 1.94, 95% CI 1.20-3.14; adjusted P=0.006) in a logistic regression model after controlling age, left atrial dimension, and the use of angiotensin-converting enzyme inhibitors and/or angiotensin receptor blockers. The T allele carriers also had increased plasma MMP-9 levels compared with CC homozygotes in both AF patients and control subjects. No relationship between R279Q polymorphism and AF was found in this cohort. CONCLUSIONS: The -1562C>T polymorphism of MMP-9 gene is significantly associated with AF risk in Chinese Han patients with hypertensive heart disease. The -1562T allele which is associated with increased expression of MMP-9 might be a genetic risk for the development of AF in this cohort.

[Detection of IFN Response of Non-Specific Effects on RNAi.].

BACKGROUND: The RNA interference (RNAi) approach is an efficient and widely used method for silencing specific genes and exploring their functions. However, numerous studies have highlighted the non-specific effects of RNAi experiments and the IFN response is one of the most common non-specific effects. In this research, we will study how to detect the IFN response in RNAi experiments. METHODS: Five different FLJ20420 siRNAs was transfected into human lung adenocarinoma cell line A549 using LipofectamineTM2000. The alternation of the interferon stimulated genes (ISGs) expression profile was analysed by real-time PCR and microarray assay. RESULTS: (1) Different FLJ20420 siRNA had different effect on silencing FLJ20420 mRNA expression in A549 cells, and FLJsiRNA-1 and -4 had the better effects to knockdown the expression of FLJ20420 (80% and 90% down-regulation, respectively); (2) In analyzed 16 major ISGs, the expression of 14 of these genes was significantly increased in A549-FLJsiRNA-1 cells (P <0.05), while only 2 of these genes had a significant change in A549-FLJ-siRNA-4 cells. These results indicated an IFN response existed in A549-FLJ-siRNA-1 cells. (3) Microarray assay indicated that the expression of 51 ISGs were increased over 2 times in A549-FLJ-siRNA-1 cells, while only the expression of 6 ISGs were over 2-time in A549-FLJ-siRNA-4 cells. These results confirmed that there was an IFN response of non-specific effects of RNAi in A549 cells transfected with FLJ-siRNA-1. CONCLUSIONS: In RNAi experiments, siRNA can not only silence the target genes, but also may activate IFN response. The analysis of the expression change of the major ISGs by real-time PCR is a useful approach to explore the IFN response in RNAi experiments.