IIIA IIIB Research Articles

Comparative Effectiveness of Intensity-Modulated Versus 3D Conformal Radiation for Stage III Non-small Cell Lung Cancer in the Medicare Population

Purpose/Objective(s): The clinical benefit of intensity-modulated (IMRT) compared to 3D conformal radiation (3D-RT) has not been wellestablished for locally advanced non-small cell lung cancer (NSCLC). We evaluated trends in use of IMRT for stage III lung NSCLC and compared survival and hospitalization outcomes. Materials/Methods: Using SEER-Medicare data, we identified use of IMRT or 3D-RT among 7061 Medicare beneficiaries diagnosed with stage III NSCLC from 2002-2009. Factors associated with use of IMRT versus 3D-RT were identified using multivariable logistic regression. Overall survival and number of hospital days within 90 days of radiation were analyzed using Cox proportional hazard and negative binomial regression models, respectively. Propensity score adjustment was used to control for clinical and demographic variables. Results: IMRT comprised an increasing proportion of conformal treatments for NSCLC, rising from 3.0% in 2002 to 26.8% in 2009. Patients treated at freestanding versus hospital-based facilities were twice as likely to receive IMRT (17.3% vs 9.5%, adj ORZ 2.0, p < 0.01). IMRT use varied by region, with higher rates in the South (12.8%, adj OR Z 1.13) and West (14.2%, adj ORZ 1.25), compared to theNortheast (9.9%, ref) andMidwest (9.1%, adjOR Z 0.88) (overall pZ 0.03) and in urban versus rural areas (12.5% vs 9.9%, adj OR Z 1.52, p < 0.01). Patients with more comorbidities were more likely to receive IMRT, 11.3% (ref) vs 11.7% (adj OR Z 1.03) vs 14.7% (adj OR Z 1.35) for modified Charlson score 0, 1, and 2+, respectively (overall pZ 0.03). We did not find a difference in IMRT use between stage IIIA and stage IIIB patients (11.8% vs 12.4%, adj OR Z 1.11, p Z 0.19). Patients receiving chemotherapy were more likely to receive IMRT (12.8% vs 10.4%, adj ORZ 1.24, p Z 0.02), though there was no difference in IMRT use among patients having surgery (11.4% vs 12.2%, adj ORZ 0.95, pZ 0.68). With propensity score adjustment, IMRTwas associatedwith greater overall survival (adj HRZ 0.91, pZ 0.03) compared to 3D-RT, though there was no difference in survival among patients receiving at least 25 fractions of radiation (adj HRZ 0.99, pZ 0.83). There was no significant difference in number of hospital days in the 90 days following radiation start (mean 5 days, adj HR Z 1.01, p Z 0.89). Conclusions: When radiation is used to treat locally advanced NSCLC, IMRT is increasingly preferred to 3D-RT. However, among patients receiving potentially curative radiation ( 25 fractions) there was no significant difference in overall survival or time spent hospitalized following treatment compared to 3D-RT. Author Disclosure: A.B. Chen: None. L. Li: None. A. Cronin: None. D. Schrag: None.

Comparative Effectiveness of Stereotactic Body Radiation Therapy Versus Surgery for Stage I Non-small Cell Lung Cancer

Purpose/Objective(s): The clinical benefit of intensity-modulated (IMRT) compared to 3D conformal radiation (3D-RT) has not been wellestablished for locally advanced non-small cell lung cancer (NSCLC). We evaluated trends in use of IMRT for stage III lung NSCLC and compared survival and hospitalization outcomes. Materials/Methods: Using SEER-Medicare data, we identified use of IMRT or 3D-RT among 7061 Medicare beneficiaries diagnosed with stage III NSCLC from 2002-2009. Factors associated with use of IMRT versus 3D-RT were identified using multivariable logistic regression. Overall survival and number of hospital days within 90 days of radiation were analyzed using Cox proportional hazard and negative binomial regression models, respectively. Propensity score adjustment was used to control for clinical and demographic variables. Results: IMRT comprised an increasing proportion of conformal treatments for NSCLC, rising from 3.0% in 2002 to 26.8% in 2009. Patients treated at freestanding versus hospital-based facilities were twice as likely to receive IMRT (17.3% vs 9.5%, adj ORZ 2.0, p < 0.01). IMRT use varied by region, with higher rates in the South (12.8%, adj OR Z 1.13) and West (14.2%, adj ORZ 1.25), compared to theNortheast (9.9%, ref) andMidwest (9.1%, adjOR Z 0.88) (overall pZ 0.03) and in urban versus rural areas (12.5% vs 9.9%, adj OR Z 1.52, p < 0.01). Patients with more comorbidities were more likely to receive IMRT, 11.3% (ref) vs 11.7% (adj OR Z 1.03) vs 14.7% (adj OR Z 1.35) for modified Charlson score 0, 1, and 2+, respectively (overall pZ 0.03). We did not find a difference in IMRT use between stage IIIA and stage IIIB patients (11.8% vs 12.4%, adj OR Z 1.11, p Z 0.19). Patients receiving chemotherapy were more likely to receive IMRT (12.8% vs 10.4%, adj ORZ 1.24, p Z 0.02), though there was no difference in IMRT use among patients having surgery (11.4% vs 12.2%, adj ORZ 0.95, pZ 0.68). With propensity score adjustment, IMRTwas associatedwith greater overall survival (adj HRZ 0.91, pZ 0.03) compared to 3D-RT, though there was no difference in survival among patients receiving at least 25 fractions of radiation (adj HRZ 0.99, pZ 0.83). There was no significant difference in number of hospital days in the 90 days following radiation start (mean 5 days, adj HR Z 1.01, p Z 0.89). Conclusions: When radiation is used to treat locally advanced NSCLC, IMRT is increasingly preferred to 3D-RT. However, among patients receiving potentially curative radiation ( 25 fractions) there was no significant difference in overall survival or time spent hospitalized following treatment compared to 3D-RT. Author Disclosure: A.B. Chen: None. L. Li: None. A. Cronin: None. D. Schrag: None.

Clinical and demographic study on open fractures caused by motorcycle traffic accidents.

OBJECTIVE: To assess socio-demographic characteristics of individuals that suffered open fractures caused by motorcycle traffic accident and evaluate infection rate in search of associated risk factors. METHODS: A retrospective study comprising 81 patients with open fractures caused by motorcycle accidents was carried out. Clinic and socio-demographic features were collected from patients' records. Comparison between infected and non-infected patients was performed to find out which variables were possibly associated to this complication. RESULTS: Patients were mostly young adults (mean 32.9 years old), of the masculine gender (83.9%), single (60.5%), from the country side (40.7%), mostly presenting tibial open fractures (48.2%). Fractures type IIIA and type IIIB were the most prevalent lesions (68,8%), and soft tissue damage graded as I and II were the most frequent (64.62%). Infection was present in 23.7% of the patients and it was associated to age higher than 40 years old (p=0.011), to time delay from trauma to first surgical procedure longer than 24 hours (p=0,012), and also to soft tissue damage extent (p=0.001). CONCLUSIONS: Patients with open fractures caused by motorcycle traffic accident were mostly young single men, coming from the state capital and presented severe tibial open fractures, which 23.7% of the cases progressed to infection. Level of Evidence III, Retrospective Comparative Study.

Effect of vaginal administration of prostaglandin E2 and/or 17beta-estradiol on luteal function and histological characteristics of the cervix in cyclic pigs.

The overall objective of this study was to examine the effect of vaginal administration of prostaglandin E2 (PGE2) and/or 17beta-estradiol (E2) on luteal function maintenance and histological properties of the porcine cervix. For this purpose, crossbred gilts were divided into three groups (n=5 per group) supplied on days 11-16 of the estrous cycle with suppositories containing: (1) placebo (Group I, Control); (2) 0.4 mg of E2 (Group II); (3) 0.4 mg of E2 and 2 mg of PGE2 (Group III). Blood samples were collected on days 11-19 of the estrous cycle to determine the concentration of progesterone (P4). Additionally, to examine local effects of the hormones applied, segments from the uterine and vaginal parts of the cervix and from the ovaries were collected post-mortem. Prolonged luteal function and extended synthesis of P4 were observed in 2 of 5 gilts receiving PGE2 and E2 simultaneously (Group III). Then, these gilts were subdivided into Group IIIA (n = 2; presence of corpora lutea on the ovaries) and Group IIIB (n = 3; lack of corpora lutea). Increased levels of plasma P4 were observed in Group IIIA on days 15-19 compared to Group IIIB and on days 16-19 compared to Group I and Group II (P < 0.05; P < 0.01; P < 0.001, respectively). In the cervix of gilts in Groups II and III, enlarged blood vessels in the lamina propria of both parts of the cervix were observed. Furthermore, in Group II the epithelium of the uterine part of the cervix was thicker (P < 0.001). Our study confirmed the proposed luteotrophic/antiluteolytic actions of E2 and PGE2 applied intravaginally. These results are significant considering that very low doses of E2 were used when compared to previous attempts. Despite the inadequate response to treatments in some of the gilts, the local effects of these hormones on the histological properties of the porcine cervix suggest that further improvements in the vaginal administration route might help to elaborate new methods for enhancing the luteal function in the pig.

ISAKOS Upper Extremity Committee Consensus Statement on the Need for Diversification of the Rockwood Classification for Acromioclavicular Joint Injuries

Optimal treatment for the unstable acromioclavicular (AC) joint remains a highly debated topic in the field of orthopaedic medicine. In particular, no consensus exists regarding treatment of grade III injuries, which are classified according to the Rockwood classification by disruption of both the coracoclavicular and AC ligaments. The ISAKOS Upper Extremity Committee has provided a more specific classification of shoulder pathologies to enhance the knowledge on and clinical approach to these injuries. We suggest the addition of grade IIIA and grade IIIB injuries to a modified Rockwood classification. Grade IIIA injuries would be defined by a stable AC joint without overriding of the clavicle on the cross-body adduction view and without significant scapular dysfunction. The unstable grade IIIB injury would be further defined by therapy-resistant scapular dysfunction and an overriding clavicle on the cross-body adduction view.

MPS IIIA and MPS IIIB: a preliminary comparison of disease trajectory, using baseline data from two independent natural history studies

MPS IIIA and MPS IIIB: a preliminary comparison of disease trajectory, using baseline data from two independent natural history studies

Is &amp;lt;sup&amp;gt;18&amp;lt;/sup&amp;gt;F-FDG-PET/CT a Valid Non-Invasive Predictor for Regression Grade after Neoadjuvant Treatment in Patients with NSCLC Stage III?

Introduction: CT alone cannot provide sufficient information referring to response after neoadjuvant therapy in a timely manner. To evaluate the role of 18F-FDG-PET after neoadjuvant chemoradiation as a valid, non-invasive predictor for early therapy response and its effect on survival as compared to histopathologic tumor response, data of 32 of 210 randomized patients with NSCLC stage IIIA/IIIB, who were treated in a prospective randomized controlled multicenter trial (LUCAS- MD), were re-evaluated. Material and Methods: For 32 patients with NSCLC stage IIIA (44%) IIIB (56%) neoadjuvant treatment consisted of two to three cycles of chemotherapy (225 mg/m2 paclitaxel and carboplatin AUC 6 d1q22) and concomitant chemoradiation (50 mg/m2 paclitaxel and carboplatin AUC 2 d1, d8, d15; 1.5 Gy b.i.d. up to 45 Gy). Documentation of involved lymph node stations as detected by 18F-FDG-PET/CT and lymph node sampling during surgery according to the IASLC lymph node mapping (2009). Evaluation of histological regression grade (RG) according to Junker et al. (2001) and correlation with 18F-FDG-PET/CT for primary tumor and each lymph node station. Calculation of disease free survival using Kaplan-Meier estimates and log rank tests. Results: Actuarial tumor specific survival for the 32 patients with concomitant chemoradiation plus chemotherapy: complete vs. incomplete metabolic remission prior to surgery after 60 months: 40% vs. 24% (p = 0.018). RG III/IIb (no/less than 10% of vital tumor cells) vs. RG IIa/I (more than 10% vital tumor cells) after 60 months: 46% vs. 15% (p = 0.006). 18/32 (56%) patients had RG III/IIb, 8/32 (25%) patients had regression grade III. 1/8 pts. with RG III were in the 18F-FDG- PET/CT false positive, 10 pts. with RG IIb (i.e. all pts. with RG IIb) were in the 18F-FDG-PET/CT false negative. One patient with RG IIa was in the 18F-FDG-PET/CT false negative. Hence, the cut-off level in detecting vital tumor cells by 18F-FDG-PET/CT after neoadjuvant chemoradiation for NSCLC is about 10%. Conclusion: Histological regression grading correlates well with metabolic remission as detected by 18F-FDG-PET. Thus, 18F-FDG-PET precedes CT in measuring the tumor response and may predict long-term therapeutic outcome in patients with stage III NSCLC. Invasive staging procedures may be avoided and patients who will not profit from resection due to insufficient downstaging after neoadjuvant treatment will be easily detected by using 18F-FDG-PET as standard imaging in workup and evaluation of treatment response.

Structure of the CRISPR Interference Complex CSM Reveals Key Similarities with Cascade

SummaryThe Clustered Regularly Interspaced Palindromic Repeats (CRISPR) system is an adaptive immune system in prokaryotes. Interference complexes encoded by CRISPR-associated (cas) genes utilize small RNAs for homology-directed detection and subsequent degradation of invading genetic elements, and they have been classified into three main types (I–III). Type III complexes share the Cas10 subunit but are subclassifed as type IIIA (CSM) and type IIIB (CMR), depending on their specificity for DNA or RNA targets, respectively. The role of CSM in limiting the spread of conjugative plasmids in Staphylococcus epidermidis was first described in 2008. Here, we report a detailed investigation of the composition and structure of the CSM complex from the archaeon Sulfolobus solfataricus, using a combination of electron microscopy, mass spectrometry, and deep sequencing. This reveals a three-dimensional model for the CSM complex that includes a helical component strikingly reminiscent of the backbone structure of the type I (Cascade) family.

Changes in visual acuity and intra-ocular pressure following bleb-related infection: the Japan Glaucoma Society Survey of Bleb-related Infection Report 2

To identify changes in visual acuity and intra-ocular pressure (IOP) 12 months after the development of bleb-related infection. Data obtained from 146 eyes of 146 patients with bleb-related infection were analyzed as a part of the Japan Glaucoma Society Survey of Bleb-related Infection. Multiple logistic regression analysis was conducted to identify factors associated with poor prognosis in visual acuity and increased IOP and for being stage III. The logMAR increased by a mean of 0.140, 0.440, 1.099 and 1.122 at 12 months postinfection for stage I, II, IIIa and IIIb infections, respectively. The logMAR was significantly worse at 6 and 12 months postinfection in stage IIIb (p = 0.002 and p = 0.003, respectively; Wilcoxon signed-rank test) and at 6 months postinfection in stage IIIa (p = 0.036). The IOP was significantly elevated following infection in both stage IIIa and stage IIIb (p = 0.028 and p = 0.008 at 6 and 12 months, respectively, for stage IIIa; p = 0.002 and p = 0.005 for stage IIIb). The multiple logistic regression analysis revealed that being stage III and positive culture were significant risk factors for poor outcome for visual acuity (Odds ratio: 9.26 and 6.29, respectively) and that being stage III was a prognostic factor for increased IOP (Odds ratio: 8.33). Pseudophakia or aphakia was significantly associated with stage III and stage IIIb infections (Odds ratio: 2.85 and 6.30). Stage III bleb-related infection causes significant visual loss and IOP elevation within 12 months after development. Therefore, preventative measures should be taken, especially in cases that are pseudophakic or aphakic.

A class of symplectic partitioned Runge–Kutta methods

This paper deals with some relevant properties of Runge–Kutta (RK) methods and symplectic partitioned Runge–Kutta (PRK) methods. First, it is shown that the arithmetic mean of a RK method and its adjoint counterpart is symmetric. Second, the symplectic adjoint method is introduced and a simple way to construct symplectic PRK methods via the symplectic adjoint method is provided. Some relevant properties of the adjoint method and the symplectic adjoint method are discussed. Third, a class of symplectic PRK methods are proposed based on Radau IA, Radau IIA and their adjoint methods. The structure of the PRK methods is similar to that of Lobatto IIIA–IIIB pairs and is of block forms. Finally, some examples of symplectic partitioned Runge–Kutta methods are presented.

Membrane Phospholipids, EML4-ALK, and Hsp90 as Novel Targets in Lung Cancer Treatment

Approximately one third of patients with non-small cell lung cancer have unresectable stage IIIA or stage IIIB disease; combined cytotoxic chemotherapy and radiation therapy delivered concurrently has been established as the standard treatment for such patients. Despite many clinical trials that tested several different radiochemotherapy combinations, it seems that a plateau of efficiencies at the acceptable risk of complications has been reached. Clinical studies indicate that the improved efficacy of radiochemotherapy is associated with the radiosensitizing effects of chemotherapy. Improvement of outcomes of this combined modality by developing novel radiosensitizers is a viable therapeutic strategy. In addition to causing cell death, ionizing radiation also induces a many-faceted signaling response, which activates numerous prosurvival pathways that lead to enhanced proliferation in the endothelial cells and increased vascularization in tumors. Radiation at doses used in the clinic activates cytoplasmic phospholipase A2, leading to increased production of arachidonic acid and lysophosphatidylcholine. The former is the initial step in the generation of eicosanoids, while the later is the initial step in the formation of lysophosphatidic acid, leading to the activation of inflammatory pathways. The echinoderm microtubule-associated protein-like 4 anaplastic lymphoma kinase (EML4-ALK) is member of the insulin superfamily of receptor tyrosine kinases. The EML4-ALK fusion gene appears unique to lung cancer and signals through extracellular signal regulated kinase and phosphoinositide 3-kinase. Heat shock protein 90 (Hsp90) is often overexpressed and present in an activated multichaperone complex in cancer cells, and it is now regarded as essential for malignant transformation and progression. In this review we focus on radiosensitizing strategies involving the targeting of membrane phospholipids, EML4-ALK, and Hsp90 with specific inhibitors and briefly discuss the combination of radiation with antivascular agents.

Introduction to the Third Edition: Diagnosis and Management of Lung Cancer, 3rd ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines

Introduction to the Third Edition: Diagnosis and Management of Lung Cancer, 3rd ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines

Parametric symplectic partitioned Runge–Kutta methods with energy-preserving properties for Hamiltonian systems

Based on W-transformation, some parametric symplectic partitioned Runge–Kutta (PRK) methods depending on a real parameter α are developed. For α=0, the corresponding methods become the usual PRK methods, including Radau IA–IA¯ and Lobatto IIIA–IIIB methods as examples. For any α≠0, the corresponding methods are symplectic and there exists a value α∗ such that energy is preserved in the numerical solution at each step. The existence of the parameter and the order of the numerical methods are discussed. Some numerical examples are presented to illustrate these results.

Systemic levels of ceruloplasmin oxidase activity in allergic asthma and allergic rhinitis

Context: The role of ceruloplasmin oxidase activity (COA) involving the interaction of oxidant and antioxidant balance in allergic diseases is still unknown.Objective: Our study was designed to examine the changes in COAs in severe persistent asthma-allergic rhinitis, new diagnosed allergic asthma-allergic rhinitis and allergic rhinitis patients.Methods: The study included 20 age- and sex-matched healthy individuals as control (Group I); Group II included 15 newly diagnosed allergic asthma-allergic rhinitis; Group III included 15 patients with severe persistent asthma-allergic rhinitis and in the fourth group there were 20 patients with allergic rhinitis. Group III was divided in two groups, severe persistent asthma-allergic rhinitis who were pre-(III-A) and post-treated (III-B) with omalizumab. Group IV was divided to two groups, pretreatment (IV-A) and posttreatment (IV-B) with specific subcutaneous immunotherapy modalities. All the posttreatment measurements were 12 months after the therapy. All the patients were assessed by the skin prick test, high sensitive C-reactive protein (hs-CRP) and COA.Results: There were significant differences between Group I and Groups III-A, III-B, IV-A and IV-B; Group II and Groups III-A, III-B, IV-A and IV-B; Group III-A and Groups III-B, IV-A and IV-B; Group III-A and Groups IV-A and IV-B; and Group IV-A and IV-B. Interestingly, there was a correlation between the hs-CRP and COA levels in Group III-A.Conclusions: Our data suggest that hs-CRP and COA levels might be an indicator of an inflammation and important in revelation of patients with allergy related diseases, especially of asthma patients.

Characterisation of the bacterial community in expressed prostatic secretions from patients with chronic prostatitis/chronic pelvic pain syndrome and infertile men: a preliminary investigation

The expressed prostatic secretions (EPSs) of men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), infertile men and normal men were subjected to microbiological study. EPSs were collected from the subjects, which included 26 normal men, 11 infertile patients and 51 CP/CPPS patients. DNA was extracted from each specimen, and the V3 regions of the 16S rRNA genes were amplified using universal bacterial primers. The results showed that the EPS 16S rRNA gene-positive rate in the CP/CPPS and infertile patients was much higher than in the normal men, but without any difference among the three patient groups. The denaturing gradient gel electrophoresis (DGGE) method was used to characterize the EPS bacterial community structure of the prostate fluid from patients with CP/CPPS or infertility issues. Principal component analysis (PCA) and partial least squares (PLS) analyses of PCR-DGGE profiles revealed that the EPS bacterial community structure differed among the three groups. Three bands were identified as the key factors responsible for the discrepancy between CP/CPPS patients and infertile patients (P<0.05). Two bands were identified as priority factors in the discrepancy of category IIIA and category IIIB prostatitis patients (P<0.05). According to this research, the ecological balance of the prostate and low urethra tract, when considered as a microenvironment, might play an important role in the maintenance of a healthy male reproductive tract.

On conjugate symplecticity of B-series integrators

The long-time integration of Hamiltonian differential equations requires special numerical methods. Symplectic integrators are an excellent choice, but there are situations (e.g., multistep schemes or energy-preserving methods), where symplecticity is not possible. It is then of interest to study whether the methods are conjugate symplectic and thus have the same long-time behaviour as symplectic methods. This question is addressed in this work for the class of B-series integrators. Algebraic criteria for conjugate symplecticity up to a certain order are presented in terms of the coefficients of the B-series. The effect of simplifying assumptions is investigated. These criteria are then applied to characterize the conjugate symplecticity of implicit Runge–Kutta methods (Lobatto IIIA and Lobatto IIIB) and of energy-preserving collocation methods.

Donor Morbidity Including Biliary Complications in Living-Donor Liver Transplantation

Because of the shortage of deceased-donor livers for transplantation, living-donor liver transplantation (LDLT) has become an indispensible treatment strategy for end-stage liver disease. The critical prerequisite for LDLT is the maximal safety of healthy donors. From June 1996 to November 2010, a total of 827 completed donor hepatectomies were performed in our center. We analyzed donor morbidity associated with LDLT. There was no donor mortality. No complications were observed in 744 (90.0%) donors, and 83 (10.0%) donors experienced complications. Wound complications were most common, occurring in 48 (5.8%) patients. According to a modified Clavien classification, grade I, grade II, grade IIIa, and grade IIIb complications were experienced in 56 (67.5%), 2 (2.4%), 15 (18.1%), and 10 (12.0%) donors, respectively. Surgical or interventional management was successful in all grade IIIa and grade IIIb donors. The incidence of biliary complications was significantly higher in younger donors. Donor morbidity did not decrease below the attained level even after time had passed. This study demonstrates the safety of donor hepatectomy. Complications were relatively minor and easily controlled. The incidence of biliary complications and donor age was inversely correlated. The procedural experience of the surgeons was not associated with the donor complication rate.

Novel Approaches of Chemoradiotherapy in Unresectable Stage IIIA and Stage IIIB Non-Small Cell Lung Cancer

Approximately one third of patients with non-small cell lung cancer have unresectable stage IIIA or stage IIIB disease, and appropriate patients are candidates for chemoradiotherapy with curative intent. The optimal treatment paradigm is currently undefined. Concurrent chemoradiotherapy, compared with sequential chemotherapy and thoracic radiation therapy (TRT), results in superior overall survival outcomes as a result of better locoregional control. Recent trials have revealed efficacy for newer chemotherapy combinations similar to that of older chemotherapy combinations with concurrent TRT and a lower rate of some toxicities. Ongoing phase III trials will determine the roles of cisplatin and pemetrexed concurrent with TRT in patients with nonsquamous histology, cetuximab, and the L-BLP25 vaccine. It is unlikely that bevacizumab will have a role in stage III disease because of its toxicity. Erlotinib, gefitinib, and crizotinib have not been evaluated in stage III patients selected based on molecular characteristics. The preliminary results of a phase III trial that compared conventionally fractionated standard-dose TRT (60 Gy) with high-dose TRT (74 Gy) revealed an inferior survival outcome among patients assigned to the high-dose arm. Hyperfractionation was investigated previously with promising results, but adoption has been limited because of logistical considerations. More recent trials have investigated hypofractionated TRT in chemoradiotherapy. Advances in tumor targeting and radiation treatment planning have made this approach more feasible and reduced the risk for normal tissue toxicity. Adaptive radiotherapy uses changes in tumor volume to adjust the TRT treatment plan during therapy, and trials using this strategy are ongoing. Ongoing trials with proton therapy will provide initial efficacy and safety data.

Podocalyxin expression in renal tissues and correlation with the number of urinary podocytes in children with Henoch-Schonlein purpura nephritis.

To analyze the podocalyxin (PCX) expression in the kidney and the number of urinary podocytes in different pathological grades of Henoch-Schonlein purpura nephritis (HSPN), and to determine whether the number of urinary podocytes reflects the renal damage in HSPN. Fifty-six children diagnosed with HSPN in our hospital were enrolled in the study and classified into 4 groups by renal pathology: grade II (IIa+IIb) (n=10), grade III (IIIa+IIIb) (n=21), grade IV (n=16), and grade V (n=9). Four kidney autopsy specimens without histomorphologic lesions and 8 urine samples from healthy children served as controls. With immunofluorescence assay, the PCX expression in 4 normal renal tissues and in the renal tissues of the 56 HSPN children was detected and quantitatively analyzed. Positive rate and the number of urinary podocytes were detected in the 8 healthy children and 56 HSPN children. In the renal tissues of the normal control group and grade II (IIa+IIb) HSPN group, the PCX expression was complete. The percentage of the PCX positive area out of the total glomerular area in the renal tissues of 2 groups had no significant difference (P>0.05). In the renal tissues of grade III (IIIa+IIIb), IV, and V HSPN groups, the PCX expression showed various degrees of loss, decreasing in turn from grade II (IIa+IIb), III (IIIa+IIIb), IV to V, with significant differences between each group (P<0.01). For HSPN with grade III (IIIa+IIIb) or higher, positive PCX expression was found in the urine, suggesting the presence of enough podocytes in the urine. The percentage of fluorescence positive area out of the total glomerular area of PCX in the renal tissues was negatively correlated with the total number of urinary podocytes (r=-0.637, P<0.01). Podocyte injury plays a certain role in the pathological progression of HSPN. The urinary detection of podocytes can reflect the degrees of pathological damage in HSPN.

A forward genetic screen to study mammalian RNA interference - essential role of RNase IIIa domain of Dicer1 in 3′ strand cleavage of dsRNA in vivo

RNA interference is a major post-transcriptional regulatory pathway in many eukaryotes. The RNase III enzyme Dicer1 processes precursor RNAs into small RNA duplexes to be loaded onto Argonaute proteins, the effector components of RNA-induced silencing complex. Biochemical studies have shown that the RNase IIIa and RNase IIIb domains of Dicer1 cleave the 3' and 5' strands of dsRNAs, respectively, although the in vivo functional significance of this activity remains unclear. Genetic screening of mammalian cells is useful for studying molecular mechanisms at the cellular level. In the present study, we conducted a novel forward genetic screen for mammalian RNA interference components using Chinese hamster ovary cells and successfully obtained several Dicer1 mutant lines. One mutant bore an intriguing Dicer1 allele in which a conserved glutamic acid in the RNase IIIa domain was substituted with a lysine. Our detailed cell biological study demonstrated that the RNase IIIa domain of Dicer1 was essential for generating small RNAs embedded in the 3' stem of exogenous hairpin-like RNAs. In the mutant cells, the expression of endogenous mature microRNAs derived from the 3' stem of pre-microRNA was repressed more severely than that from the 5' stem. Moreover, appropriate processing and loading of small RNAs were required for the dissociation of Argonaute 2 from Dicer1. The data obtained in the present study demonstrate that this screening method represents a promising strategy for the identification of unknown components of mammalian RNA interference pathways and the study of the biological significance of these components at the cellular level.