[Recurrence and metastasis of pheochromocytoma/paraganglioma after tumor resection and clinical characteristics analysis].

Objective: The purpose of this study is to investigate the incidence of recurrence or metastasis of pheochromocytoma/paraganglioma (PPGL) patients after primary tumor resection, and to compare the differences of clinical indicators between patients with or without recurrence or metastasis. Methods: This study is a retrospective study. All 157 patients were followed up after tumor resection in Peking Union Medical College Hospital from 2008 to 2016. We obtained the basic information [age of diagnosis, gender, height, weight and body mass index (BMI)], the onset status of PPGL (age of onset, course of disease, family history, tumor location, whether the tumor is bilateral or multiple, and preoperative blood pressure), clinical and pathological features of PPGL tumor (tumor size, whether it could adhere, invade or infiltrate during operation, whether the tumor capsule is smooth and complete on the postoperative pathological diagnosis, whether there is infiltration growth and cystic necrosis on tumor pathology and Ki-67 index), and laboratory examination results [24 hours urinary norepinephrine (NE), epinephrine (E), dopamine (DA) before operation]. According to the outpatient or telephone follow-up, the postoperative incidences of recurrence and metastasis were explored, and the basic information, status of onset, clinical and pathological characteristics of tumors, and laboratory test results of patients were compared. Results: A total of 157 patients, 69 males and 88 females, were with an average age of (42.4±13.4) years old. There were 103 patients with PCC and 54 with PGL. The average follow-up time was (9.5±2.0) years. Of the 103 patients with PCC, 13 (12.6%) had postoperative recurrence and 9 (8.7%) had distant metastasis. Compared with the patients without recurrence and metastasis, the onset age of the 13 patients with recurrence was younger [(27.3±15.7) years vs (39.3±12.2) years, P=0.003], the course of disease was longer [48.0 (23.0, 141.0) months vs 12.0 (4.0, 60.0) months, P=0.010]. The tumor size of 9 patients with distant metastasis was larger [8.0 (6.1, 12.8) cm vs 5.0 (4.0, 7.0) cm, P=0.027]. Of the 54 patients with PGL, 8 (14.8%) had postoperative recurrence and 5 (9.3%) had distant metastasis. Compared with the patients without recurrence and metastasis, the course of disease of the 8 patients with recurrence was longer [90.0 (36.3, 165.0) months vs 24.0 (8.0, 72.0) months, P=0.009], and the proportion of primary tumors with multiple lesions was higher (4/8 vs 4.4%, P=0.003). The preoperative diastolic blood pressure was higher in 5 patients with distant metastasis [(146.0±32.1) mmHg vs (120.6±25.3) mmHg, P=0.043] (1 mmHg=0.133 kPa), and the proportion of primary tumors with multiple lesions was higher (2/4 vs 4.4%, P=0.029). Conclusion: PPGL patients are prone to have recurrence or metastasis. PPGL patients with postoperative recurrence or distant metastasis had younger onset age, longer course of disease, larger tumor size and higher proportion of multiple lesions.

YBX1 Protects against Apoptosis Induced by Oxygen-Glucose Deprivation/Reoxygenation in PC12 Cells via Activation of the AKT/GSK3ß Pathway.

Reperfusion therapies for ischaemic stroke can induce secondary injury accompanied by neuronal death. The Y-box binding protein 1 (YBX1), an oncoprotein, is critical for regulating tumour cell proliferation and apoptosis. Thus, we wanted to know whether YBX1 could regulate neuronal cell apoptosis caused by cerebral ischaemia/reperfusion (I/R). We established a model of cerebral I/R-induced injury in vitro by oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and determined YBX1 expression using Western blot. Next, the effect of YBX1 on the apoptosis and viability of OGD/R-treated PC12 cells was evaluated by flow cytometry, MTT assay, and Western blot. Besides, the release of lactate dehydrogenase (LDH) and the activity of catalase (CAT) and superoxide dismutase (SOD) were detected to evaluate oxidative stress of PC12 cells induced by OGD/R. The regulatory roles of YBX1 in the AKT/GSK3ß pathway were examined by Western blot. As a result, OGD/R treatment down-regulated YBX1 expression in PC12 cells. YBX1 over-expression attenuated the growth inhibition and apoptosis of PC12 cells induced by OGD/R. Besides, the increase of LDH release and the decrease of SOD and CAT activities caused by OGD/R were reversed by YBX1 over-expression. Moreover, YBX1 over-expression could activate the AKT/GSK3ß pathway in OGD/ R-treated PC12 cells. Therefore, YBX1 could protect against OGD/R-induced injury in PC12 cells through activating the AKT/GSK3ß signalling pathway, and thus YBX1 has the potential to become a therapeutic target for cerebral I/R-induced injury.

[Task-related functional connectivity of anterior cingutate in opiate drug addicts during physical detoxification: a task fMRI study].

Objective: To explore the brain areas which have abnormal functional connectivity with anterior cingutate in opiate drug dependence during physical detoxification using a task-related functional magnetic resonance imaging(fMRI), and to find out the role of anterior cingutate dysfunction in the relapse of opiate drug addicts during physical detoxification. Methods: From January to December 2016, eighteen participants of opiate drug addicts during physical detoxification who completed in Drug Rehabilitation Center of Anhui Province, and eighteen healthy controls recruited performed a cue-elicited craving task in a MRI scanner while signal data were collected. Two regions of interest were the right anterior cingutate and the left anterior cingutate, then the linear correlation between the whole brain and the anterior cingutates was calculated to find out the abnormal functional connectivity of the anterior cingutates. Results: Contrasted experimental group with the healthy controls, the functional connectivity of bilateral fusiform gyrus, caudate nucleus, and the anterior cingutates was increased in the opiate drug addicts during physical detoxification group (P<0.05),and the functional connectivity between anterior cingutates and polus temporalis, hippocampi, Middle frontal gyrus of orbit, Supplementary motor area, dorsolateral superior frontal gyrus was decreased(P<0.05). Conclusion: The anterior cingutates dysfunction of functional connectivity in a cue-elicited craving task may play a important role in the relapse of opiate drug addicts during physical detoxification.

[Comparison of the accuracy of neuronavigation and linear measurement in the positioning of parasagittal meningioma].

Objective: To explore the accuracy of preoperative positioning of neuronavigation and linear measurement localization in the parasagittal meningioma. Methods: Thirty-eight patients with parasagittal meningioma who underwent neurosurgery in the First Affiliated Hospital of Bengbu Medical College from January 2016 to April 2018 were preoperatively positioned with neuronavigation localization and linear measurement localization. The actual position of the tumor was observed during operation, and the accuracy of the two methods was compared. The time taken by the neural navigation positioning and the linear measurement positioning method was recorded, and the difference between the two methods was compared. Results: All cases were treated with neuronavigation and linear measurement, and the tumors were completely exposed after localization. Compared with the actual tumor center position measured during surgery, the average error distance between 38 nerve navigation and actual tumor center position was (2.7±1.9) mm, and the average error distance between linear measurement and actual tumor center position was (3.2±1.3) mm. The difference was not statistically significant, P value=0.207. Neuronavigation includes booting, data import, registration, positioning, etc., which takes an average of (22.3±2.3) minutes. The linear positioning method included image data measurement and localization, and the average time was(1.7±0.3) minutes. The difference between the two groups was statistically significant, and the P value was less than 0.001. Conclusion: The linear measurement localization method and the neuronavigation localization method have good accuracy for preoperative localization of the parasagittal meningioma.

Recognizing 50 Years of Innovative Newborn Screening in North Carolina.

Newborn screening is a critical, preventive health program for early identification of disorders in newborns. Early detection, diagnosis, and treatment of certain genetic or metabolic disorders can significantly reduce death, disease, and associated disabilities. North Carolina has been instrumental in the development of innovative technologies used in newborn screening programs.

[The curative effect analysis of unilateral subfrontal combined with interhemispheric approach on the microsurgery of large anterior skull base meningioma].

Objective: To explore the surgical technique and curative effect of microsurgical resection of large anterior skull base meningioma via unilateral subfrontal and interhemispheric approach. Method: The clinical data of 14 patients with large anterior skull base meningioma who received surgical treatment in the Department of neurosurgery in First Affiliated Hospital of Bengbu Medical College from April 2015 to September 2017 were analysed retrospectively.Of 9 cases were olfactory groove meningioma and 5 cases were tuberculum sellae meningioma.The microsurgical resection approach of all the patients was the unilateral subfrontal combined with interhemispheric approach. Results: According to the Simpson classification of meningioma resection, 13 cases(92.8%) were Simpson grade Ⅰand Ⅱ, 1 case was grade Ⅲ.The clinical symptoms were improved in 12 cases (85.7%), and no improvement in 2 cases.Fourteen patients had postoperative complications in 2 cases (14.2%), and all of them were no cerebrospinal fluid leakage and death. Conclusion: Microsurgical removal of large anterior skull base meningioma through unilateral frontal and interhemispheric approach is safe and effective, providing a new way for surgical treatment of large anterior skull base meningioma. It had a positive significance for the prognosis of patients.

[Application of modified brain protection double-lumen drainage tube in external drainage of chronic subdural hematoma].

Objective: To compare the clinical effects of modified double-lumen drainage tube and traditional silicone tube in external drainage of chronic subdural hematoma. Methods: Selected 49 patients suffering from chronic subdural hematoma hospitalized in the Department of Neurosurgery, the First Affiliated Hospital of Bengbu Medical College between July 2016 and June 2018 who underwent external drainage, and divided them randomly into experimental groups (using modified brain protection double lumen drainage tube in 23 cases) and control group (using traditional silicone drainage tube in 26 cases). Comparison of postoperative hematoma residual volume, the tube indwelling time, the numbers of drainage tube contact with brain tissue and arachnoid membrane, the numbers of infections, the numbers of postoperative epilepsy cases, and changes in Modified Rankin Scale scores between the two groups. Results: At the time of extubation, the residual amount of hematoma in the experimental group was (13±7) ml, and that in the control group was (17±8) ml. There was no significantly statistical difference in the residual amount of hematoma between the experimental group and the control group (P>0.05). The tube indwelling time of the experimental group was (2.0±0.9) days, and that of the control group was (2.7±0.8) days. The difference between the experimental group and the control group in the drainage tube indwelling time was statistically significant (P<0.05). No cases of drainage tube contact with brain tissue and/or arachnoid membrane appeared in the experimental group, and 7 cases of drainage tube contact with brain tissue or arachnoid membrane appeared in the control group. No infection occurred in both groups. No cases of epilepsy occurred in preoperative, and there were 0 cases of epilepsy in the experimental group and 1 case in the control group in postoperative. There were significantly statistical differences in each of the two groups in the modified Rankin scale before and after surgery (P<0.001), there was no significant difference in postoperative Modified Rankin Scale scores between the two groups. Conclusion: The modified brain protection double-lumen drainage tube has good drainage effect in the external drainage of chronic subdural hematoma, and the short tube retention time, causing fewer complications. It is a safe and effective tool for treating chronic subdural hematoma, and it is worthy of clinical promotion.

Factors influencing suicidal ideation among Chinese patients with stomach cancer: qualitative study.

AIM: The aim of this qualitative descriptive study was to explore the influencing factors of suicidal ideation in Chinese patients diagnosed with stomach cancer. INTRODUCTION: More recently, links have been made between suicide and cancer. For inpatients with cancer, nurses are at the front line to cope with suicidal issues in clinical settings. METHOD: This study was part of a larger study that investigated psychological strains and related factors associated with suicidal ideation. Thirty-two patients with stomach cancer participated in a face-to-face interview. Thematic analysis was used to analyse the qualitative data. FINDINGS: Four themes were identified as protective factors against suicidal ideation: suicidal denial and fears of stigmatization; positive coping skills; family connectedness; and desire to live well and longer. DISCUSSION: Consistent with previous research, these four protective factors may encourage participants to think more positively about life and cancer. CONCLUSION: Our findings suggest four protective factors for targeted suicide prevention, which may play an important role in future suicide-prevention programmes for Chinese patients diagnosed with stomach cancer. IMPLICATIONS FOR NURSING: Nurses may help reduce patients' risk for suicidal ideation more effectively by discussing beliefs, family and meaning in life perspectives, as well as offering health education about positive coping skills. IMPLICATIONS FOR HEALTHY POLICY: Clinical nurses, hospitals and the government should work collaboratively with each other. It is suggested that Chinese national guidelines for working with the high-risk suicide populations should be considered within a future mental health working plan.

[Curative effect analysis of two surgical methods for removal of pituitary adenoma via endonasal transsphenoidal approach].

Objective: To compare the efficacy of endoscopic endonasal transsphenoidal surgery and microsurgery for pituitary adenomas. Methods: One hundred and ten patients with pituitary adenoma who were treated by single nostril transsphenoidal surgery in the department of neurosurgery of the First Affiliated Hospital of Bengbu Medical College from June 2014 to December 2016 were enrolled.These cases were randomly divided into endoscopic group 53 cases (including 36 cases of functional pituitary adenoma and 17 cases of non-functional pituitary adenoma) and microscope group 57 cases (including 34 cases of pituitary adenoma and 23 cases of non-functional pituitary adenomas), with no significant difference in preoperative clinical data about gender, age, tumor size and endocrine function (P>0.05). The total tumor resection rate, postoperative complication rate and the rate of functional tumor hormone levels were compared between the two groups. Results: The total resection rate of tumor in patients with endoscope and microscope group were 75.5% (40/53) and 70.2% (40/57) without statistically significant difference (P>0.05). The total resection rate and hormone level decline effective rate of functional pituitary adenomas in neuroendoscope group (91.7%, 33/36; 83.3%, 30/36) were higher than those in microscope group (70.6%, 24/34; 61.8%, 21/34) with statistically significant difference (P<0.05). Another side, there was no statistically significant difference in total resection of non-functional pituitary adenomas between endoscope group (41.2%, 7/17) and microscope group (69.6%, 16/23) (P>0.05). The total incidence of postoperative complications in endoscopic group (9.4%, 5/53) was lower than that in the microscope group (24.6%, 14/57), with statistically significant difference (P<0.05), in which the rate of postoperative complications of functional pituitary adenomas and nonfunctional pituitary adenoma in endoscopy group (8.3%, 3/36; 11.8%, 2/17) and in microscope (20.6%, 7/34; 30.4%, 7/23) had no statistical significance (P>0.05). Conclusion: Pituitary adenoma resection rate in two ways had no obvious difference, but the advantages of endoscopic surgery for functional pituitary adenomas were obvious.The incidence of postoperative complications of endoscopic surgery was lower than that of microscopic surgery, but there was no difference between functional and non-functional tumors.

[Curative effect analysis of 15 cases of cervical intra-and extra-spinal tumors by microsurgical resection].

Objective: To explore the curative effect of microsurgical treatment of cervical intra- and extra-spinal tumors. Methods: The clinical data of 15 patients with cervical intra-and extra-spinal tumors in Department of Neurosurgery, the First Affiliated Hospital of Bengbu Medical College from January 2013 to February 2017 were retrospectively analyzed. According to Shaw's tumor stage: 6 cases in stage Ⅰ, 5 cases in stage Ⅱ, 3 cases in stage Ⅲ, and 1 case in stage Ⅳ. About the surgical methods: 9 cases were treated via posterior median semi-laminectomy approach, of which were internal fixation in 2 cases, and via posterior median approach spinous process and vertebral plate complex orthotopic replantation to spinal canal plasty in 3 cases, other 3 cases were operated via anterior cervical approach, including internal fixation in 1 case. After the surgery, vertebral canal MRI was adopted to evaluate the tumor, and CT three-dimensional reconstruction or X-ray of the spine was used for assessing the stability of spinal column, and Frankel grading standard was employed to evaluate the recovery of spinal cord function. Results: The tumors were totally resected in the first stage with the help of a microscope. Histological types of the tumors included schwannoma in 14 cases, gangliocytoma in 1 case. There was no leakage of cerebrospinal fluid and infection of incisional wound after operations. After the surgery, Frankel grading standard was adopted to assess the recovery of the spinal cord function, and the results showed grade D in 4 patients and grade E in 11 patients. All patients were followed-up for 1 to 24 months with an average of 13 months without tumor recurrence and spinal instability. Conclusion: Appropriate surgical approach should be selected to treat the cervical intra- and extra-spinal tumors, totally resection of the tumor could be achieved under the microscope, and the corresponding vertebral reconstruction and spinal fixation should be used to maintain spinal stability if necessary.

[Research on the application of "H shaped" single-tube double-lumen drainage tube in the treatment of chronic subdural hematoma].

OBJECTIVE: To explore the effect of chronic subdural hematoma external drainage surgery using self-made "H shaped" flush type single-tube double-lumen drainage tube. METHODS: There were 56 cases chosen from the First Affiliated Hospital of Bengbu Medical College between Jan 2013 and Aug 2015. These patients with unilateral chronic subdural hematoma requiring surgery to place drilling external drainage catheter were randomly divided into group A (21 cases, using self-made single-tube double lumen "H shaped" drainage tube) and group B (35 cases, traditional silicone drainage tube), then the residual liquid volume after drainage on the first day, the days that the tube stay in body and the residual fluid volume after removing the tube were compared between the two groups. RESULTS: The residual liquid volume after drainage on the first day in group A was (23±15)ml, in group B was (31±15)ml. The days that the tube stay in body in group A was (2.7±1.0)d, in group B was (3.3±1.1)d, the two groups had statistical differences (P<0.05). The residual fluid volume after removing the tube in group A was (13±7) ml, in group B was (16±8)ml, but the data in these two groups had no significantly statistical differences (P>0.05). CONCLUSION: The effect of self-made "H shaped" flush type single-tube double-lumen drainage tube in the drainage of chronic subdural hematoma drainage is good, with short tube stay in the body; therefore, it is a safe and effective way to treat chronic subdural hematoma, and is worthy of clinical application.

[Determination of relative elements of hard metal in workplace air and urine by inductive coupled plama].

Objective: To establish a rapid detection method regarding the air conditions of workplace and the workers' urine included Tungsten, Cobalt, Nickel, Titanium, Cadmium, Manganese, Lead and its compounds based on inductively coupled plasma mass spectrometry (ICP-MS) . Methods: The experiment adopts ICP-MS to deter-mine those metals in workshop air and workers urine, evaluate the detection's limitation, the precision and accuracy of the method. Using the membrane filter and urine freeze - dried metal standard material to verify this method. Results: Each element of correlation coefficient was greater than 0.999. The recovery rate of air samples was 91.6%~104.6%, within-batch RSD precision was 1.41%~3.50%, between-run precision was 1.28%~4.31%, urine samples recovery rate was 93.0%~102.6%, within - batch RSD precision was 1.25%~3.56%, between - run precision was 1.58%~4.67%, According to the method every element was within the scope of the standard reference, it was also showed that the established method is accurate and reliable. Conclusion: ICP-MS is an effective and feasible method to detect the workshop air and the workers' urine which included Tungsten, Cobalt, Nickel, Titanium, Cadmium, Manganese, Lead and its compounds.

Acetylpuerarin reduces inflammation and improves memory function in a rat model of Alzheimer's disease induced by Abeta1-42.

This study was performed to determine if acetylpuerarin (compound N-2211) could reduce amyloid-beta1-42 (Abeta1-42) induced learning and memory deficits and to examine its anti-neuroinflammatory effects in a rat model. Forty Wistar rats were randomly divided into four groups (n = 10 each): control, model (Abeta1-42 injected), low-dose and high-dose acetylpuerarin groups. The acetylpuerarin groups received peritoneal acetylpuerarin every day for 12 days after 2 weeks of Abeta1-42 (5 microg/1 microl) intrahippocampal injections. The Morris water maze (MWM) was used to assess rats' learning and memory abilities. Immunohistochemistry was used to assess expression levels of ionized calcium-binding adaptor molecule (Ibal), protein kinase C delta (PKCdelta), IkappaB kinase beta (IKKbeta), and inducible nitric oxide synthase (iNOS) in hippocampus. After Abeta1-42 injection, the learning and memory abilities of rats were reduced, and acetylpuerarin treatment ameliorated the observed deficits. Abeta1-42 injection resulted in microglia transforming from resting microglia into an activated state, but this was reduced by acetylpuerarin treatment. Furthermore, hippocampal expression of PKCdelta, IKKbeta, and iNOS increased following Abeta1-42 treatment, and acetylpuerarin could suppressed the levels of PKCdelta, iNOS, and IKKbeta. Acetylpuerarin improves learning and memory functions in Abeta1-42 induced rat models. These effects may be due to anti-neuroinflammatory effects.

International Consensus on Minimum Preclinical Testing Requirements for the Development of Innovative Therapies For Children and Adolescents with Cancer.

Cancer remains the leading cause of disease-related death in children. For the many children who experience relapses of their malignant solid tumors, usually after very intensive first-line therapy, curative treatment options are scarce. Preclinical drug testing to identify promising treatment elements that match the molecular make-up of the tumor is hampered by the fact that (i) molecular genetic data on pediatric solid tumors from relapsed patients and thus our understanding of tumor evolution and therapy resistance are very limited to date and (ii) for many of the high-risk entities, no appropriate and molecularly well-characterized patient-derived models and/or genetic mouse models are currently available. However, recent regulatory changes enacted by the European Medicines Agency (class waiver changes) and the maturation of the RACE for Children act with the FDA, will require a significant increase in preclinical pediatric cancer research and clinical development must occur. We detail the outcome of a pediatric cancer international multistakeholder meeting whose output aims at defining an international consensus on minimum preclinical testing requirements for the development of innovative therapies for children and adolescents with cancer. Recommendations based on the experience of the NCI funded PPTP/C (www.ncipptc.org) and the EU funded ITCC-P4 public private partnership (www.itccp4.eu) are provided for the use of cell-based and mouse models for pediatric solid malignancies, as well as guidance on the scope and content of preclinical proof-of-concept data packages to inform clinical development dependent on clinical urgency. These recommendations can serve as a minimal guidance necessary to jumpstart preclinical pediatric research globally.

EpCAM in carcinogenesis: the good, the bad or the ugly.

The epithelial cell adhesion molecule (EpCAM) is a membrane glycoprotein that is highly expressed on most carcinomas and therefore of potential use as a diagnostic and prognostic marker for a variety of carcinomas. Interestingly, EpCAM is explored as target in antibody-based therapies. Recently, EpCAM has been identified as an additional marker of cancer-initiating cells. In this review, we describe the controversial biological role of EpCAM with the focus on carcinogenesis: as an adhesion molecule, EpCAM mediates homophilic adhesion interactions, which in turn might prevent metastasis. On the other hand, EpCAM abrogates E-cadherin mediated cell-cell adhesion thereby promoting metastasis. Also, upon cleavage of EpCAM, the intracellular domain functions as a part of a transcriptional complex inducing c-myc and cyclin A and E. In line with these seemingly controversial roles, EpCAM overexpression has been associated with both decreased and increased survival of patients. Similarly, either induction or downregulation of EpCAM expression lowers the oncogenic potential depending on the cell type. As epigenetic dysregulation underlies aberrant EpCAM expression, we propose epigenetic editing as a novel approach to investigate the biological role of EpCAM, expanding the options for EpCAM as a therapeutic target in cancer.

Sustained performance by red and white muscle fibres from the dogfish Scyliorhinus canicula.

The mechanical performance of red and white muscle fibres from dogfish was compared during a long series of contractions with sinusoidal movement or under isometric conditions at 12 degrees C (normal in vivo temperature). Power output was measured during sinusoidal movement at 0.75 Hz and peak-to-peak amplitude about 12% L(0). Tetanus duty cycle was 33% (0.44 s) at phase -8% (first stimulus at 0.107 s before shortening started). Initially, the red fibres produced only about one third as much power as the white fibres, 6.57+/-0.63 W kg(-1) wet mass (mean +/- s.e.m.) and 18.3+/-2.3, respectively. Red fibres were better at sustaining power output; it declined rapidly to about 60% of its initial value and then remained relatively steady for up to 450 cycles of movement. Force during shortening declined, but force during stretch did not increase: force always relaxed to a low value before stretch started. By contrast, net power output by white fibres declined rapidly to zero within about 50 cycles. Two changes contributed: decline in force during shortening and an increase in force during stretch because relaxation became progressively less complete during the series of contractions. In isometric series (0.44 s stimulation every 1.33 s, cycle frequency 0.75 Hz), red and white fibres sustained peak isometric force similarly; in the 50th cycle force was 59+/-3% and 56+/-4% of initial values. The time required for force to relax to 10% of its maximum value decreased during the series for red fibres and increased for white fibres.

Endonucleases induced TRAIL-insensitive apoptosis in ovarian carcinoma cells.

TRAIL induced apoptosis of tumor cells is currently entering phase II clinical settings, despite the fact that not all tumor types are sensitive to TRAIL. TRAIL resistance in ovarian carcinomas can be caused by a blockade upstream of the caspase 3 signaling cascade. We explored the ability of restriction endonucleases to directly digest DNA in vivo, thereby circumventing the caspase cascade. For this purpose, we delivered enzymatically active endonucleases via the cationic amphiphilic lipid SAINT-18((R)):DOPE to both TRAIL-sensitive and insensitive ovarian carcinoma cells (OVCAR and SKOV-3, respectively). Functional nuclear localization after delivery of various endonucleases (BfiI, PvuII and NucA) was indicated by confocal microscopy and genomic cleavage analysis. For PvuII, analysis of mitochondrial damage demonstrated extensive apoptosis both in SKOV-3 and OVCAR. This study clearly demonstrates that cellular delivery of restriction endonucleases holds promise to serve as a novel therapeutic tool for the treatment of resistant ovarian carcinomas.

The rates of second lung cancers and the survival of surgically-resected second primary lung cancers in patients undergoing resection of an initial primary lung cancer.

BACKGROUND: The Lung Cancer Screening Trial demonstrated improved overall survival (OS) and lung cancer specific survival (LCSS), likely due to finding early-stage NSCLC. The purpose of our investigation is to evaluate whether long-term surveillance strategies (4+ years after surgical resection of the initial lung cancer(1LC)) would be beneficial in NSCLC patients by assessing the rates of second lung cancers(2LC) and the OS/LCSS in patients undergoing definitive surgery in 1LC as compared to 2LC (>48 months after 1LC) populations. METHODS: SEER13/18 database was reviewed for patients during 1998-2013. Log-rank tests were used to determine the OS/LCSS differences between the 1LC and 2LC in the entire surgical group(EG) and in those having an early-stage resectable tumors (ESR, tumors <4 cm, node negative). Joinpoint analysis was used to determine rates of second cancers 4-10 year after 1LC using SEER-9 during years 1985-2014. RESULTS: The rate of 2LCs was significantly less than all other second cancers until 2001 when the incidence of 2LCs increased sharply and became significantly greater than all other second cancers in females starting in year 2005 and in men starting in year 2010. OS/LCSS, adjusted for propensity score by using inverse probability weighting, demonstrated similar OS, but worse LCSS for 2LCs in the EG, but similar OS/LCSSs in the ESR group. CONCLUSION: Because the rate of 2LCs are increasing and because the OS/LCSS of the 1LC and 2LC are similar in early-stage lesions, we feel that continued surveillance of patients in order to find early-stage disease may be beneficial.

Targeted delivery of a designed sTRAIL mutant results in superior apoptotic activity towards EGFR-positive tumor cells.

Previously, we have shown that epidermal growth factor receptor (EGFR)-selective delivery of soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL), by genetic fusion to antibody fragment scFv425, enhances the tumor-selective pro-apoptotic activity of sTRAIL. Insight into the respective contribution of the agonistic receptors TRAIL-R1 and TRAIL-R2 to TRAIL-induced apoptosis may provide a rational approach to further optimize TRAIL-based therapy. Recently, this issue has been investigated using sTRAIL mutants designed to selectively bind to either receptor. However, the relative contribution of the respective TRAIL receptors, in particular TRAIL-R1, in TRAIL signaling is still unresolved. Here, we fused scFv425 to designed sTRAIL mutant sTRAILmR1-5, reported to selectively activate TRAIL-R1, and investigated the therapeutic apoptotic activity of this novel fusion protein. EGFR-specific binding of scFv425:sTRAILmR1-5 potently induced apoptosis, which was superior to the apoptotic activity of scFv425:sTRAIL-wt and a nontargeted MOCK-scFv:sTRAILmR1-5. During cotreatment with cisplatin or the histone deacetylase inhibitor valproic acid, scFv425:sTRAILmR1-5 retained its superior pro-apoptotic activity compared to scFv425:sTRAIL-wt. However, in catching-type Enzyme-Linked ImmunoSorbent Assays with TRAIL-R1:Fc and TRAIL-R2:Fc, scFv425:sTRAILmR1-5 was found to not only bind to TRAIL-R1 but also to TRAIL-R2. Binding to TRAIL-R2 also had functional consequences because the apoptotic activity of scFv425:sTRAILmR1-5 was strongly inhibited by a TRAIL-R2 blocking monoclonal antibody. Moreover, scFv425:sTRAILmR1-5 retained apoptotic activity upon selective knockdown of TRAIL-R1 using small inhibitory RNA. Collectively, these data indicate that both agonistic TRAIL receptors are functionally involved in TRAIL signaling by scFv425:sTRAILmR1-5 in solid tumor cells. Moreover, the superior target cell-restricted apoptotic activity of scFv425:sTRAILmR1-5 indicates its therapeutic potential for EGFR-positive solid tumors.

Reversal of blindness in animal models of leber congenital amaurosis using optimized AAV2-mediated gene transfer.

We evaluated the safety and efficacy of an optimized adeno-associated virus (AAV; AAV2.RPE65) in animal models of the RPE65 form of Leber congenital amaurosis (LCA). Protein expression was optimized by addition of a modified Kozak sequence at the translational start site of hRPE65. Modifications in AAV production and delivery included use of a long stuffer sequence to prevent reverse packaging from the AAV inverted-terminal repeats, and co-injection with a surfactant. The latter allows consistent and predictable delivery of a given dose of vector. We observed improved electroretinograms (ERGs) and visual acuity in Rpe65 mutant mice. This has not been reported previously using AAV2 vectors. Subretinal delivery of 8.25 x 10(10) vector genomes in affected dogs was well tolerated both locally and systemically, and treated animals showed improved visual behavior and pupillary responses, and reduced nystagmus within 2 weeks of injection. ERG responses confirmed the reversal of visual deficit. Immunohistochemistry confirmed transduction of retinal pigment epithelium cells and there was minimal toxicity to the retina as judged by histopathologic analysis. The data demonstrate that AAV2.RPE65 delivers the RPE65 transgene efficiently and quickly to the appropriate target cells in vivo in animal models. This vector holds great promise for treatment of LCA due to RPE65 mutations.