Can hemifacial spasm lead to glaucomatous changes?

PURPOSE: To evaluate the association between hemifacial spasm (HFS) patients and glaucoma as a function of the Botox dosage required. METHODS: A retrospective review of clinical documents and procedure records. RESULTS: Information of 76 consecutive patients (58 females) with HFS who received Botox treatment were reviewed. The age at onset of HFS was 66±11 (32-85) years, and all manifested unilaterally. Ten (13%, 95% confidence interval: 6.5-22.9%) patients were diagnosed with glaucoma, including 8 primary open-angle glaucoma (POAG) (4 unilateral and ipsilateral to the HFS), and 2 bilateral chronic angle-closure glaucoma (CACG). Nine of the 10 patients developed glaucoma after the onset of the HFS. The Botox dosage was significantly higher among those diagnosed with glaucoma (31+/8 vs. 26+/7units, P<0.05). There was a positive relationship between the presenting intraocular pressure (IOP) and the Botox dosage required (R=0.31, P=0.0116). However, there was a weak relationship between the Botox dosage required and the vertical cup to disc ratio (R=0.076, P=0.525). The presenting IOP of the HFS-affected eyes in those diagnosed with glaucoma was higher than those without glaucoma (19±3.5 vs. 13±3.2mmHg, P=<0.05). The presenting IOP between the HFS-affected and unaffected eyes was similar (16±4.8 vs. 15+/4.6mmHg, P=0.430). Smoking status, history of diabetes mellitus, hypertension, hyperlipidemia and obstructive sleep apnea were not different between HFS patients with or without glaucoma. CONCLUSIONS: Hemifacial spasm patients with glaucoma were associated with a higher Botox dosage. We found a positive relationship between the Botox dosage required and the presenting IOP. Whether hemifacial spasm can result in fluctuation of IOP, eventually causing glaucomatous damage, remains to be studied further.

Downregulation of linc00961 contributes to promote proliferation and inhibit apoptosis of vascular smooth muscle cell by sponging miR-367 in patients with coronary heart disease.

OBJECTIVE: Atherosclerosis is one of the most important risk factors for coronary heart disease (CHD), and growing evidence has shown that long non-coding RNAs (lncRNAs) can serve as prospective markers for atherosclerosis. In this study, we mainly focused on the potential roles of linc00961 in CHD patients. PATIENTS AND METHODS: qRT-PCR was used to detect the expressions of linc00961 and miR-367 in CHD patients and ApoE-/-mice, and the correlations were analyzed. Then, HA-VAMC was respectively treated with 5 inflammatory factors and hypoxia conditions to explore the factors that affect linc00961 levels. Furthermore, the linc00961 overexpression lentivirus (LV-linc00961) and linc00961 downregulation lentivirus (LV-sh linc00961) were purchased and transfected into human vascular smooth muscle cells (VSMCs). CCK8 assay was carried out to measure the cell proliferation of VSMC, and the levels of Cyclin D1, Bcl-2, Bax, and cleaved caspase-3 were detected by RT-PCR and Western blot. Moreover, the Luciferase assay was performed to explore the binding site of linc00961 and miR-367. Finally, the miR-367 inhibitor was transfected into LV-sh linc00961 VSMCs to confirm the linc00961 functions via miR-367. RESULTS: We found that linc00961 was significantly decreased in patients with CHD and ApoE-/-mice. Additionally, linc00961 was reduced in VSMCs at the conditions of hypoxia and C-reactive protein (CRP). Most importantly, the overexpression of linc00961 significantly inhibited the VSMCs proliferation, repressed the levels of Cyclin D1 and Bcl-2, and increased the levels of Bax and cleaved caspase-3. However, the downregulation of linc00961 promoted VSMCs proliferation, increased the levels of Cyclin D1 and Bcl-2, and repressed the levels of Bax and cleaved caspase-3. We also found that miR-367 was downregulated following the upregulation of linc00961, while it was upregulated following the downregulation of linc00961. The Luciferase gene reporter assay indicated that linc00961 could directly bind with miR-367 in VSMCs. Finally, we found that linc00961 could inhibit proliferation and promote apoptosis of VSMCs via binding with miR-367. CONCLUSIONS: According to the results, our study revealed that linc00961 was significantly decreased in patients with CHD and ApoE-/-mice. Furthermore, our findings firstly uncovered that linc00961 was reduced by hypoxia and CRP in VSMCs. The downregulation of linc00961 contributed to promote proliferation and inhibit apoptosis of VSMCs by sponging miR-367 in CHD patients, which might provide a potential target for treating atherosclerosis.

High-quality thulium iron garnet films with tunable perpendicular magnetic anisotropy by off-axis sputtering - correlation between magnetic properties and film strain.

Thulium iron garnet (TmIG) films with perpendicular magnetic anisotropy (PMA) were grown on gadolinium gallium garnet (GGG) (111) substrates by off-axis sputtering. High-resolution synchrotron radiation X-ray diffraction studies and spherical aberration-corrected scanning transmission electron microscope (Cs-corrected STEM) images showed the excellent crystallinity of the films and their sharp interface with GGG. Damping constant of TmIG thin film was determined to be 0.0133 by frequency-dependent ferromagnetic resonance (FMR) measurements. The saturation magnetization (Ms) and the coercive field (Hc) were obtained systematically as a function of the longitudinal distance (L) between the sputtering target and the substrate. A 170% enhancement of PMA field (H⊥) was achieved by tuning the film composition to increase the tensile strain. Moreover, current-induced magnetization switching on a Pt/TmIG structure was demonstrated with an ultra-low critical current density (jc) of 2.5 × 106 A/cm2, an order of magnitude smaller than the previously reported value. We were able to tune Ms, Hc and H⊥ to obtain an ultra-low jc of switching the magnetization, showing the great potential of sputtered TmIG films for spintronics.

Effects of individual and partner factors on anxiety and depression in Taiwanese prostate cancer patients: A longitudinal study.

Studies exploring the mediating and predictive factors of anxiety and depression for prostate cancer patients in Eastern countries are scant. Guided by the transactional model of stress and coping, this study determined the predictors and mediators of anxiety and depression in prostate cancer patients. The participants comprised 115 prostate cancer patients and 91 partners. The patients and partners completed questionnaires regarding physical symptoms, disease appraisals, coping behaviours, anxiety and depression in the period before confirmation of treatment decisions and 1, 3, 6 and 12 months after treatment. The results revealed that partner anxiety engendered a stressful situation and aggravated patient anxiety. Patients' threat appraisals and affective-oriented coping behaviours mediated the relationships between their anxiety levels and those of their partners. The patients' most recent prostate-specific antigen (PSA) levels and hormonal symptoms were key predictors of their anxiety and depression levels. The patients' harm appraisals mediated the relationships between their most recent PSA levels and hormonal symptoms and depression. Their threat appraisals and affective-oriented coping behaviours mediated the relationships between their hormonal symptoms and anxiety and depression. To manage those key factors, reframing, appraising disease and improving coping behaviours may reduce anxiety and depression levels in prostate cancer patients.

[Lung function influenced the prognosis of cardiac surgery in patients with chronic obstructive pulmonary disease].

Objective: To investigate the value of lung function for the prognosis of cardiac surgery in patients with chronic obstructive pulmonary disease(COPD). Methods: A retrospective analysis was conducted on the hospitalized patients with coronary heart disease or valvular heart disease in Beijing Anzhen hospital during Janury 2013 to December 2015. The relationship between lung function and extubation time, time of staying in ICU, second time tracheal incubation, tracheotomy and mortality rate were analyzed. Results: There were 355 patients without surgery in a total 1 729 of patients, of which 65 (18.31%)suffered from severe pulmonary dysfunction. There were 242 patients with obstructive ventilation dysfunction, 75 with mild, 127 with moderate, and 40 with severe and very severe obstructive ventilation dysfunction. There were significant differences in the rates of planned extubation and late extubation between patients with normal lung function (81.6% and 10.7%) and those with abnormal lung function(74.4% and 12.8%). In patients with different GOLD classification (Ⅰ, Ⅱ, and Ⅲ-Ⅳ), there were significant differences in the rates of early extubation 14.7%, 14.2% and 5%, planned extubation (80%, 74% and 65%) and late extubation (5.3%, 11.8% and 30%). By comparison with patients without COPD, the extubation time of COPD patients increased by 20.3%, the time of staying in ICU prolonged by 14.2%, and the mortality rate increased to 180%(P<0.05). However, there was no significant difference in the rate of second time tracheal incubation or tracheotomy. Conclusion: There was a close relationship between GOLD classification and prognosis of cardiac surgery patients with COPD. Cardiac surgery needs to be cautious because of increase in incidence of postoperative adverse events in patients with severe pulmonary dysfunction.

Comparison of immediate breast reconstruction after mastectomy and mastectomy alone for breast cancer: A meta-analysis.

BACKGROUND: The purpose of this study was to compare patient outcomes between immediate breast reconstruction (IBR) after mastectomy and mastectomy alone. METHODS: We conducted a comprehensive literature search of PUBMED, EMBASE, Web of Science, and Cochrane Library. The primary outcomes evaluated in this review were overall survival, disease-free survival and local recurrence. Secondary outcome was the incidence of surgical site infection. All data were analyzed using Review Manager 5.3. RESULTS: Thirty-one studies, involving of 139,894 participants were included in this paper. Pooled data demonstrated that women who had IBR after mastectomy were more likely to experience surgical site infection than those treated with mastectomy alone (risk ratios 1.51, 95% CI: 1.22-1.87; p = 0.0001). There were no significant differences in overall survival (hazard ratios 0.92, 95% CI: 0.80-1.06; p = 0.25) and disease-free survival (hazard ratios 0.96, 95% CI: 0.84-1.10; p = 0.54) between IBR after mastectomy and mastectomy alone. No significant difference was found in local recurrence between two groups (risk ratios 0.92, 95% CI: 0.75-1.13; p = 0.41). CONCLUSIONS: Our study demonstrates that IBR after mastectomy does not affect the overall survival and disease-free survival of breast cancer. Besides, no evidence shows that IBR after mastectomy increases the frequency of local recurrence.

Effects of ovarian ablation or suppression in premenopausal breast cancer: A meta-analysis of randomized controlled trials.

BACKGROUND: The effect of ovarian ablation or suppression (OAS) in premenopausal women with breast cancer is controversial. The overall survival (OS), disease-free survival (DFS) and adverse event of OAS versus no OAS were compared. METHODS: A literature review of EMBASE, Web of Science, PUBMED, and Cochrane Library was conducted. The hazard ratio (HR) and 95% confidence interval (CI) for OS and DFS, as well as risk ratio (RR) and 95% CI for adverse events were evaluated. I-squared statistic (I2) represents heterogeneity. RESULTS: Twenty-nine studies with a total of 21,249 women were included. In premenopausal women aged 40 years or younger, there were significant differences in OS (HR 0.78, 95% CI: 0.66-0.94, P=0.008, I2 = 0%) and DFS (HR 0.84, 95% CI: 0.73-0.97, P=0.02, I2 = 0%) between OAS and no OAS. In advanced stage breast cancer, a significant difference was found in OS (HR 0.76, 95% CI: 0.60-0.96, P=0.02, I2 = 0%). Patients treated with OAS had more chances to have hot flushes (RR 1.91, 95% CI: 1.62-2.26, P < 0.01, I2 = 0%) and vaginal dryness (RR 1.19, 95% CI: 1.08-1.31, P=0.0003, I2 = 0%). No significant difference in depression (RR 1.28, 95% CI: 0.94-1.74, P=0.12, I2 = 0%). CONCLUSIONS: The study shows that OAS plays a beneficial role in premenopausal women aged 40 years or younger and advanced stage breast cancer. However, OAS is associated with increase in hot flushes and vaginal dryness.

Infiltrating immune cells and gene mutations in pancreatic ductal adenocarcinoma.

BACKGROUND: The aim of this study was to assess the immune profile within the microenvironment of pancreatic ductal adenocarcinoma (PDAC), and to investigate the prognostic value of intratumoral infiltrating immune/inflammatory cells (IICs) in patients after surgery. METHODS: Eighteen phenotypic markers representing 11 types of IIC and the protein products of genes TP53, CDKN2A/p16 and SMAD4/DPC4 were assessed by immunohistochemistry of specimens from patients with pancreatic cancer. The expression of IICs and the mutational status of the genes were correlated with tumour recurrence and survival, and results were validated in an independent cohort. RESULTS: CD15+ neutrophils, CD20+ B cells and CD206+ tumour-associated macrophages were seen frequently in tumours, and their presence was associated with reduced survival in a cohort of 79 patients. Expression of CD4+ T helper cells, CD8+ cytotoxic T lymphocytes and CD117+ mast cells was associated with a favourable prognosis. A weighted Cox regression recurrence-predictive model was constructed that showed good correlation of IICs and gene mutations. A combination of CD15, CD206, CD117 and Smad4 expression was independently associated with overall (hazard ratio (HR) 3·63, 95 per cent c.i. 2·18 to 6·04; P < 0·001) and recurrence-free (HR 2·93, 1·81 to 4·75; P < 0·001) survival. These findings were validated in an independent cohort (151 patients) and in 54 tissue samples obtained by preoperative endoscopic ultrasound-guided fine-needle aspiration. CONCLUSION: PDAC has a unique immunosuppressive phenotype that is associated with characteristic gene mutations, disease recurrence and survival after pancreatectomy. Surgical relevance The immune microenvironment plays a critical role in the development of pancreatic ductal adenocarcinoma (PDAC). PDAC is associated with mutations in major driver genes, including KRAS, TP53, CDKN2A/p16 and SMAD4/DPC4. This study shows that the microenvironment of PDAC has a unique immunosuppressive phenotype, which may be driven by oncogene mutations. Patients with PDAC with a highly immunosuppressive profile tended to have poor postoperative survival. A model including three intratumoral infiltrating immune markers (CD15+, CD206+ and CD117+) and a SMAD4 mutation can be used to predict recurrence and survival in patients after surgery for PDAC.

Axillary lymph node dissection versus sentinel lymph node biopsy alone for early breast cancer with sentinel node metastasis: A meta-analysis.

BACKGROUND: In early breast cancer patients with sentinel node metastasis, the effect of axillary lymph node dissection (ALND) is controversial. The purpose of this study is to compare the safety and efficacy of sentinel lymph node biopsy (SLNB) alone versus ALND in patients with early breast cancer and sentinel node metastasis. METHODS: We searched PubMed, Embase, Web of Science, and Cochrane Library databases from 1965 to February 2014. All data were analyzed using Review Manager Software 5.2. RESULTS: 12 studies, which included 130,575 patients from five randomized controlled trials and seven observational studies, met our inclusion criteria. 26,870 early breast cancer patients underwent SLNB alone and 103,705 underwent ALND. Patients underwent ALND had more paresthesia (risk ratio [RR] 0.26, 95% confidence interval [CI] 0.20-0.33; p < 0.01) and lymphedema (RR 0.28, 95% CI 0.20-0.41; p < 0.01) than those had SLNB alone. There were no significant differences in overall survival (hazard ratio [HR] 0.95, 95% CI 0.85-1.06; p = 0.35), disease-free survival (HR 1.00, 95% CI 0.98-1.02, p = 0.96), and locoregional recurrence (RR 0.92, 95% CI 0.59-1.44; p = 0.73). CONCLUSION: Current evidence indicates that axillary dissection may be omitted in early breast cancer patients with sentinel lymph metastasis.

Androgen deprivation increases the risk of fracture in prostate cancer patients: a population-based study in Chinese patients.

UNLABELLED: Androgen deprivation therapy (ADT) or orchiectomy is associated with an increased risk of osteoporosis or fracture. In this nationwide database analysis, we found that ADT or orchiectomy increased the risk of fracture in Chinese patients with prostate cancer. However, the magnitude of increase is seemingly not as large as that in Western populations. INTRODUCTION: ADT using gonadotropin-releasing hormone (GnRH) agonists or orchiectomy is associated with an increased risk of osteoporosis or fracture. To investigate the effects of ADT duration or orchiectomy on any type of fracture in Asian patients with prostate cancer, we conducted this retrospective analysis using a nationwide database in Taiwan. METHODS: We included 17,359 subjects who were newly diagnosed with prostate cancer between January 1, 1998, and December 31, 2007. The risk of first fracture was our primary endpoint. RESULTS: The rates of fracture from 12 months after prostate cancer diagnosis until the last follow-up date were 8.7 % for all patients, 7.1 % for patients who did not receive ADT or orchiectomy, 9.8 % for patients who received ADT, and 14.4 % for patients who received orchiectomy with or without ADT (P < 0.0001). In a Cox proportional hazard analysis, the relative risk of fracture increased steadily with the number of doses of GnRH agonists received during the first year after cancer diagnosis and with dose density. A significant hazard ratio was observed in patients who received at least nine doses within 1 year after diagnosis and in those whose dose density exceeded two doses per year. Age greater than or equal to 65 years was associated with a significantly lower risk of fracture. CONCLUSION: ADT or orchiectomy increases the risk of fracture in Chinese patients with prostate cancer. However, the magnitude of this increase is seemingly not as large as that in Western populations.

Antitumor effects of oncolytic adenovirus armed with PSA-IZ-CD40L fusion gene against prostate cancer.

Advanced prostate cancer (PC) still remains incurable. Novel immunogene therapy shows promise as treatment strategy that can target both localized and metastasized PC. In this study, we have developed a PC-specific oncolytic adenovirus (Ad-PL-PPT-E1A) armed with fusion gene of prostate-specific antigen and CD40 ligand, and aimed to evaluate its therapeutic effect in vitro and in vivo. After they were rescued in human embryonic kidney 293 cells, we confirmed that Ad-PL-PPT-E1A could mediate the expression of E1A efficiently and produce abundant progeny viruses in PC cells in vitro. Our data showed that Ad-PL-PPT-E1A induced apoptosis and resulted in specific oncolytic toxicity in PC cells, which was detected by Annexin-V staining and crystal violet, respectively. After stimulation with lysates, immune phenotypes and cytokines expression of human dendritic cells was detected by flow cytometry and real-time polymerase chain reaction, respectively. And, the results showed that the lysate of Ad-PL-PPT-E1A-infected LNCaP cells upregulated the expression of CD80, CD83, CD86 and mRNA level of interleukin-6 (IL-6), IL-12, IL-23 and tumor necrosis factor-α significantly. In established PC3M cell-xenografted mouse models, Ad-PL-PPT-E1A treatment improved the survival and suppressed the tumor growth obviously. In conclusion, Ad-PL-PPT-E1A exhibited enhanced antitumor activity is a promising approach for gene therapy of advanced PC.

Cavin-1 is essential for the tumor-promoting effect of caveolin-1 and enhances its prognostic potency in pancreatic cancer.

Caveolin-1 exhibits a stage-dependent, functional fluctuation during pancreatic cancer development, but the underlying mechanisms remain unclear. Here, we report that cavin-1, a structural protein of caveolae, modulates the oncogenic function of caveolin-1 and cooperates with caveolin-1 to enhance pancreatic cancer aggressiveness. Cavin-1 expression is associated with caveolin-1 in pancreatic cancer tissue samples and cell lines, and predicts the metastatic potential of pancreatic cancer. Interactome analyses further revealed the physical interaction of cavin-1 and caveolin-1 and their colocalization in pancreatic cancer cells. Cavin-1 stabilizes caveolin-1 expression or activity by inhibiting its internalization and subsequent lysosomal degradation. More in-depth functional experiments showed that caveolin-1-enhanced aggressiveness of pancreatic cancer cells is dependent on the presence of cavin-1. In contrast, cavin-1 depletion inhibited the invasion and metastasis of pancreatic cancer cells, which could not be restored by caveolin-1-rescue construct. Tissue microarray analyses in two independent clinic cohorts also supported the augment of cavin-1 on the prognostic potency of caveolin-1, and showed that combination of cavin-1 with caveolin-1 predicted worse survival in pancreatic cancer patients. Of note, the phenotypes because of cavin-1 could not be achieved by other cavins such as cavin-2, and the tumor-promoting role of cavin-1 in pancreatic cancer was found to be largely dependent on caveolin-1 expression, which highlights the critical role of cavin-1/caveoin-1 in pancreatic cancer progression, and suggests that the interruption of cavin-1/caveolin-1 interaction is a promising therapeutic strategy for pancreatic cancer.

Neural stem cells harvested from live brains by antibody-conjugated magnetic nanoparticles.

It stems from the magnetism: The extraction of stem/progenitor cells from the brain of live animals is possible using antibodies conjugated to magnetic nanoparticles (Ab-MNPs). The Ab-MNPs are introduced to a rat's brain with a superfine micro-syringe. The stem cells attach to the Ab-MNPs and are magnetically isolated and removed. They can develop into neurospheres and differentiate into different types of cells outside the subject body. The rat remains alive and healthy.

Long-term survival after hand-assisted laparoscopic approach of primary retroperitoneal mucinous cystadenocarcinoma in male: case report and review of literature.

BACKGROUND: Primary retroperitoneal mucinous cystadenocarcinoma (PRMC) is a rare disease and mostly occurs in females, and there are only three male cases described in the literatures without long-term follow-up. CASE REPORT: A 59-year-old male presented with a left retroperitoneal cystic mass (7.5 ´ 7 ´ 3 cm) that upwardly displaced the left kidney and caused abdominal discomfort. The tumor was totally excised by the hand-assisted laparoscopic method without complications or recurrence in a follow-up period of 79 months. The etiology from coelomic metaplasia of peritoneal epithelium was proved by a spectrum of diverse cells (benign, borderline malignant, and malignant cells) during pathological examination. RESULTS: This is the fourth male case of PRMC in the world with a favorable outcome after hand-assisted laparoscopic excision, and this is also distinct by the longest follow-up period in this disease entity. CONCLUSIONS: Because of its low-malignant potential and recurrence rate, surgical excision is still the best choice of treatment, but the least invasion method should be adopted in front.

Complicated features in a young child with influenza B virus pneumonia and co-infection with Stenotrophomonas maltophilia.

A 3.5-year-old child with influenza B virus pneumonia developed pneumomediastinum and subcutaneous emphysema on the 3rd day of illness. Bronchoscopy demonstrated obstruction of the left main bronchus by mucopurulent sputum. Culture of the broncho-alveolar lavage yielded Stenotrophomonas maltophilia. After the respiratory complications resolved (11 days), the patient developed neurological symptoms and was diagnosed as acute disseminated encephalomyelitis (ADEM). Stenotrophomonas maltophilia was probably a factor in the development of pneumomediastinum. To our knowledge, this is the first case report of influenza virus infection with Stenotrophomonas maltophilia co-infection associated with spontaneous pneumomediastinum.

Efficiency of infiltration with bupivacain after modified radical mastectomy.

BACKGROUND: Pre-incisional infiltration of anaesthetic is proven to reduce postoperative pain in breast cancer surgery. However, studies of post-incisional infiltration for modified radical mastectomy are rare. The purpose of this study was to investigate whether post-incisional infiltration with bupivacaine provides improved postoperative pain relief and a cost-effective benefit. METHODS: This is a retrospective study. Between January 2006 and May 2008, 139 patients who received modified radical mastectomy were recruited to participate in the study. Patients receiving local infiltration received bupivacaine (0.5% bupivacaine, 5 ml diluted to 10 ml with distilled water) injected into the dermis surrounding the incision after wound suture. Pain intensity was evaluated using a Visual Analogue Scale (VAS) score and measurement of the required doses of meperidine and acetaminophen. The pain score was recorded every eight hours for three days. RESULTS: All patients were female. Seventy-two patients received local infiltration with bupivacaine after wound suture and 67 patients did not. There were no significant differences between the two patient groups in age, body weight and height, length of general anaesthesia and operative time. Hospital stay was significantly shorter for patients receiving local infiltration of bupivacaine. The VAS score was higher up to 16 hours post-surgery for patients who did not receive local infiltration. Meperidine and acetaminophen consumption was less for patients who received local infiltration (P = 0.010). CONCLUSION: Post-incisional wound infiltration with bupivacaine can relieve pain during the first 16 hours after surgery and shorten hospital stay, and it provides a cost-effective benefit.

Comparison of epidermal growth factor receptor mutations between primary and corresponding metastatic tumors in tyrosine kinase inhibitor-naive non-small-cell lung cancer.

BACKGROUND: Mutations of the epidermal growth factor receptor (EGFR) gene in non-small-cell lung cancer (NSCLC) patients predict the patients who will respond to EGFR tyrosine kinase inhibitor (TKI) treatment. A recent study has suggested that 33% of NSCLC showed primary tumor/metastasis discordance of EGFR expression by immunohistochemistry analysis. We intended to find out whether the EGFR mutations of primary lung cancers are concordant to that of corresponding metastatic tumors. MATERIALS AND METHODS: We analyzed EGFR exons 18-21 from paired primary and metastatic tumors in 67 lung cancer patients who had not received TKI before tissues were sampled. RESULTS: Using the direct sequencing method, 9 of 18 (50%) patients with EGFR mutation-positive primary lung tumors had lost the mutations in metastases. For 26 patients who were EGFR mutation positive in the metastatic tumors, 17 (65%) were negative in the primary tumors. We analyzed these paired tissues with discrepant EGFR mutations by the Scorpion Amplified Refractory Mutation System assay. Finally, the discordant rate reached 27% (18 of 67 cases). CONCLUSION: EGFR mutations in primary lung tumors do not always reflect the same situation in metastases. Analysis of EGFR mutations in the primary lung tumor would be inadequate for planning the use of TKI for advanced NSCLC.

Sphingosine kinase 1 gene transfer reduces postoperative peritoneal adhesion in an experimental model.

BACKGROUND: Recovery of the surgically damaged mesothelial cell layer is a major process in reducing postoperative peritoneal adhesions. Sphingosine kinase (SPK) 1 is a signalling molecule involved in the regulation of proliferation and migration of various cell types. This study determined the effect of SPK-1 gene transfer on the recovery of damaged mesothelial cells and on peritoneal adhesion formation after surgery. METHODS: Rat mesothelial cells were isolated and characterized by their expression of cytokeratin and vimentin. Their migration was determined by scratch wound motility assay. Cellular SPK-1 activity was measured by [gamma-32P]adenosine 5'-triphosphate incorporation. Wistar rats underwent laparotomy with subsequent caecum or uterine horn abrasion. Rats were randomized to either SPK-1 gene (Ad-SPK-1) transfer or control groups. The animals were killed 14 days after operation and peritoneal adhesions were graded. RESULTS: Adenovirus-mediated SPK-1 gene transfer increased the cellular SPK-1 activity of mesothelial cells, leading to enhanced migration. Median adhesion scores were significantly lower in the Ad-SPK-1 group than in controls in both rat caecum (0.98 versus 2.60; P < 0.001) and rat uterine horn (0.28 versus 1.83; P < 0.001) models. CONCLUSION: Adenovirus-mediated SPK-1 gene transfer promotes recovery of the surgically damaged mesothelial cell layer and prevents postoperative peritoneal adhesion formation.

A simplified system for generating oncolytic adenovirus vector carrying one or two transgenes.

Oncolytic adenoviruses, also called conditionally replicating adenoviruses (CRADs), have been widely applied in cancer gene therapy. However, the construction of CRADs is still time-consuming. In this study, we attempted to establish a simplified method of generating CRADs based on AdEasy system. A novel plasmid pTE-TPE-GM was constructed, containing sequentially positioned promoter of telomerase reverse transcriptase (TERTp), coding sequence of E1A gene, promoter of E1B gene, granulocyte-macrophage colony-stimulating factor (GM-CSF) gene, internal ribosome entry site sequence and coding sequence of E1B55K gene. The CRAD-generating system reported here include three plasmids: pTE-TPE-GM, pShuttle-CMV and AdEasy-1, one Escherichia coli strain BJ5183, and the packaging cell line 293. Using this system, an oncolytic adenovirus carrying B7-1 (CD80) and GM-CSF genes was successfully constructed and designated as Ad-CD80-TPE-GM. The expression of GM-CSF increased more than 9000 times in tumor cell lines infected by Ad-CD80-TPE-GM at a multiplicity of infection (MOI) of 5, compared with the cells infected by replication-defective control virus. Similarly, the expression of CD80 also increased 9-140 times. Ad-CD80-TPE-GM selectively replicates in TERT-positive tumor cells, and the progeny viruses can reach up to 375 infection units (IU) per cell. In vitro study showed that the Ad-CD80-TPE-GM induced an obvious oncolytic effect at MOI of 0.1, and killed about 80% TERT-positive tumor cells within 7 days at an MOI of 1. The antitumor effect of this vector was also investigated in Hep2 xenograft model of nude mice, and the tumor inhibition rate reached 74% at day 30 after the administration with a total dose of 1 x 10(9) IU Ad-CD80-TPE-GM. Intratumoral injection of Ad-CD80-TPE-GM slightly induced neutralizing antibody against the oncolytic adenovirus in nude mice, which might contribute to the virus clearance in vivo. In conclusion, we successfully constructed an oncolytic CRAD carrying GM-CSF and CD80 gene. More importantly, this system can be modified to generate novel transcriptionally regulated CRADs with different tissue-specific promoters or transgenes.