The current study aim was to identify predictors of pathologic complete response (pCR) following neoadjuvant therapy.
From 2000-2007, 518 breast cancer patients received neoadjuvant therapy. Data were compared using Chi-square, Fisher’s exact test, and MANOVA, where appropriate.
Of 518 breast cancer patients receiving neoadjuvant therapy, 81 (16%) had a pCR [77 of 456 (17%) chemotherapy; 4 of 62 (6%) endocrine therapy; p<0.05]. Four factors were associated with pCR: higher tumor grade (p=0.015), lack of estrogen and progesterone receptor (ER/PR) expression (p<0.0001), HER-2/neu amplification (p=0.025), and negative lymph node status (p<0.0001). On multivariate analysis, ER/PR negativity, HER-2/neu amplification, and negative lymph node status were found to significantly correlate with pCR.
Patients with ER/PR negative and Her-2/neu amplified breast cancer phenotypes are more likely to experience a pCR to neoadjuvant therapy. While a pCR is more frequently observed following neoadjuvant chemotherapy, it is rare following neoadjuvant endocrine therapy.
Breast cancer; Neoadjuvant therapy; Pathologic response
Margin status is a significant risk factor for local recurrence. We sought to examine whether the method of tumor localization predicted the margin status and the need for re-excision for both non-palpable and palpable breast cancer.
We identified 358 consecutive breast cancer patients who were treated with breast-conserving therapy (BCT) from 1999–2006. Data included patient and tumor characteristics, method of localization (needle versus palpation), and pathologic outcomes. Descriptive statistics were utilized for data summary and data were compared using Chi-square.
Of 358 patients undergoing BCT, 234 (65%) underwent needle localization for a non-palpable tumor and 124 (35%) underwent a palpation-guided procedure. Patients undergoing palpation-guided procedures were younger and had larger tumors at a more advanced pathologic stage of disease than those undergoing needle localization procedures (p<0.05 for each). Patient race, tumor grade, presence of lymphovascular invasion, biomarker profile, and nodal status were not significantly different between the two groups (p>0.05). Overall, 137 (38%) patients had one or more positive margins; 90 of 234 (38%) who had a needle localization procedure and 47 of 124 (38%) who had a palpation-guided procedure (p>0.05). The number of margins affected did not differ significantly between the two groups.
Although patients with palpable breast cancer had larger tumors than those with non-palpable breast cancer, the incidence and number of positive margins was similar to those who had needle localization for non-palpable tumors. Improved methods of localization are needed to reduce the rate of positive margins and the need for re-excision.
Breast Cancer; Margin Status; Needle Localization
We investigated factors associated with positive margins following mastectomy and the impact on outcomes.
We identified 240 patients with stage I-III invasive breast cancer who underwent mastectomy from 1999-2009. Data included patient and tumor characteristics, pathologic margin assessment, and outcomes. Margin positivity was defined as the presence of in situ or invasive malignancy present at any margin. Descriptive statistics were utilized for data summary and were compared using Chi-square.
Of the 240 patients, 132 (55%) had a simple mastectomy with sentinel lymph node biopsy and 108 (45%) had a modified radical mastectomy. Overall, 21 (9%) patients had positive margins, including 12 (57%) with one positive margin, 3 (14%) with two positive margins, and 6 (29%) with three or more positive margins. The most commonly affected margin was the deep margin (48% of patients). Eight (38%) of the 21 patients received adjuvant chest wall irradiation. There were no differences between patients who had a positive margin versus those who did not with respect to patient age, race, percentage of in situ component, tumor size, tumor grade, lymphovascular invasion, or immunostain profile (p>0.05 for all). None of the patients with positive margins experienced a local recurrence.
Positive margins following mastectomy occurred in nearly 10% of our patients. No specific patient or tumor characteristics predicted a risk for having a positive margin. Despite the finding that only approximately 40% of patients received adjuvant radiation in the setting of a positive margin, no local recurrences have been observed.
Breast cancer; Margin status; Mastectomy; Radiation therapy
Donor specific HLA antibodies significantly lower allograft survival, but as yet there are no satisfactory therapies for prevention of antibody-mediated rejection. Intracapillary macrophage infiltration is a hallmark of antibody-mediated rejection, and macrophages are important in both acute and chronic rejection. The purpose of this study was to investigate the Fc-independent effect of HLA I antibodies on endothelial cell activation, leading to monocyte recruitment. We used an in vitro model to assess monocyte binding to endothelial cells in response to HLA I antibodies. We confirmed our results in a mouse model of antibody-mediated rejection, in which B6.RAG1-/- recipients of BALB/c cardiac allografts were passively transferred with donor specific MHC I antibodies. Our findings demonstrate that HLA I antibodies rapidly increase intracellular calcium and endothelial presentation of P-selectin, which supports monocyte binding. In the experimental model, donor specific MHC I antibodies significantly increased macrophage accumulation in the allograft. Concurrent administration of rPSGL-1-Ig abolished antibody-induced monocyte infiltration in the allograft, but had little effect on antibody-induced endothelial injury. Our data suggest that antagonism of P-selectin may ameliorate accumulation of macrophages in the allograft during antibody-mediated rejection.
Endothelial cells; HLA antibody; signal transduction; monocytes; P-selectin
rabies; rabies virus; viruses; zoonoses; China; Henan Province
Maintenance and release of seed dormancy is regulated by plant hormones; their levels and seed sensitivity being the critical factors. This study reports transcriptional regulation of brassinosteroids (BR), ethylene (ET), cytokinin (CK) and salicylic acid (SA) related wheat genes by after-ripening, a period of dry storage that decays dormancy. Changes in the expression of hormonal genes due to seed after-ripening did not occur in the anhydrobiotic state but rather in the hydrated state. After-ripening induced dormancy decay appears to be associated with imbibition mediated increase in the synthesis and signalling of BR, via transcriptional activation of de-etiolated2, dwarf4 and brassinosteroid signaling kinase, and repression of brassinosteroid insensitive 2. Our analysis is also suggestive of the significance of increased ET production, as reflected by enhanced transcription of 1-aminocyclopropane-1-carboxylic acid oxidase in after-ripened seeds, and tight regulation of seed response to ET in regulating dormancy decay. Differential transcriptions of lonely guy, zeatin O-glucosyltransferases and cytokinin oxidases, and pseudo-response regulator between dormant and after-ripened seeds implicate CK in the regulation of seed dormancy in wheat. Our analysis also reflects the association of dormancy decay in wheat with seed SA level and NPR independent SA signaling that appear to be regulated transcriptionally by phenylalanine ammonia lyase, and whirly and suppressor of npr1 inducible1 genes, respectively. Co-expression clustering of the hormonal genes implies the significance of synergistic and antagonistic interaction between the different plant hormones in regulating wheat seed dormancy. These results contribute to further our understanding of the molecular features controlling seed dormancy in wheat.
Optic nerve tortuosity is often reported in children with neurofibromatosis type 1 (NF1). We describe quantitative and subjective criteria to determine the degree of optic nerve tortuosity in individuals with NF1.
To employ quantitative and subjective criteria to assess optic nerve tortuosity in individuals with NF1.
Materials and methods
A retrospective study over a period of 8 years was performed on children with NF1, with and without optic pathway glioma, compared to children without NF1. A tortuosity index was computed for the optic nerve in each subject using a high resolution 3D T1-weighted magnetization-prepared rapid gradient echo sequence, which was averaged and compared across groups.
The tortuosity index for subjects with NF1, regardless of an optic pathway glioma, was greater than those without NF1. There was no difference in the tortuosity index between NF1 subjects with optic pathway glioma and NF1 subjects without optic pathway glioma. There was also no correlation between subjective measures of tortuosity and the quantitative scoring (tortuosity index), or between the degree of tortuosity and subject age or gender.
Individuals with NF1 have increased optic nerve tortuosity relative to unaffected individuals. Quantitative tortuosity index is a superior measure to subjective assessment in the evaluation of optic nerve tortuosity in children with NF1.
NF1; optic glioma; MRI; brain tumor; cancer predisposition syndrome
De novo cancers are a growing problem that has become one of the leading causes of late mortality after liver transplantation. The incidences and risk factors varied among literatures and fewer concerned the Eastern population.
The aim of this study was to examine the incidence and clinical features of de novo cancers after liver transplantation in a single Chinese center.
569 patients who received liver transplantation and survived for more than 3 months in a single Chinese center were retrospectively reviewed.
A total of 18 de novo cancers were diagnosed in 17 recipients (13 male and 4 female) after a mean of 41±26 months, with an overall incidence of 3.2%, which was lower than that in Western people. Of these, 8 (3.32%) cases were from 241 recipients with malignant liver diseases before transplant, while 10 (3.05%) cases were from 328 recipients with benign diseases. The incidence rates were comparable, p = 0.86. Furthermore, 2 cases developed in 1 year, 5 cases in 3 years and 11 cases over 3 years. The most frequent cancers developed after liver transplantation were similar to those in the general Chinese population but had much higher incidence rates.
Liver transplant recipients were at increased risk for developing de novo cancers. The incidence rates and pattern of de novo cancers in Chinese population are different from Western people due to racial and social factors. Pre-transplant malignant condition had no relationship to de novo cancer. Exact risk factors need further studies.
ZnO materials with a range of different morphologies have been successfully synthesized via a simple double-solvothermal method in the presence of glycine. The morphologies of the products can be controlled from superstructures to microrods by adjusting the amount of water in the EtOH/H2O system. Photoluminescence (PL) studies reveal that the more amount of water was used, the stronger PL relative intensity of the green emission is, but the weaker ultraviolet emission. This might be attributed to the more defects of the products when the more water was used. The catalytic studies show that all the samples have good abilities to decrease decomposition temperature around 300°C and the decomposition temperature lowers with the increase of the relative intensity of ZnO green emission.
Single-crystalline hyperbranched nanostructures of iron hydroxyl phosphate Fe5(PO4)4(OH)3·2H2O (giniite) with orthorhombic phase were synthesized through a simple route. They have a well-defined dendrite fractal structure with a pronounced trunk and highly ordered branches. The toxicity test shows that the hyperbranched nanostructures have good biocompatibility and low toxicity level, which makes them have application potentials in life science. The study herein demonstrated that the obtained hyperbranched giniite nanostructures show highly selective capture of phosphopeptides and could be used as a kind of promising nanomaterial for the specific capture of phosphopeptides from complex tryptic digests with the detection of MALDI-TOF mass spectrometry.
We investigated the patients and microbiological risk factors that predispose to the development of primary breast abscesses and subsequent recurrence.
Patients with a primary breast abscess requiring surgical therapy between January 1, 2000 and December 31, 2006 were reviewed. Recurrent breast abscess was defined by the need for repeated drainage within 6 months. Patient characteristics were compared to the general population and between groups.
A total of 89 patients with a primary breast abscess were identified; 12 (14%) were lactational and 77 (86%) were nonlactational. None of the lactational abscesses recurred, whereas 43 (57%) of the nonlactational abscesses did so (P < 0.01). Compared to the general population, patients with a primary breast abscess were predominantly African American (64% vs. 12%), had higher rates of obesity (body mass index > 30: 43% vs. 22%), and were tobacco smokers (45% vs, 23%) (P < 0.01 for all). The only factor significantly associated with recurrence in the multivariate logistic regression analysis was tobacco smoking (P = 0.003). Compared to patients who did not have a recurrence, patients with recurrent breast abscesses had a higher incidence of mixed bacteria (20.5% vs. 8.9%), anaerobes (4.5% vs. 0%), and Proteus (9.1% vs. 4.4%) but lower incidence of Staphylococcus (4.6% vs. 24.4%) (P < 0.05 for each).
Risk factors for developing a primary breast abscess include African American race, obesity, and tobacco smoking. Patients with recurrent breast abscesses are more likely to be smokers and have mixed bacterial and anaerobic infections. Broader antibiotic coverage should be considered for the higher risk groups.
Our study aims were to investigate breast cancer patients with micrometastases or isolated tumor cells (ITCs) in sentinel lymph nodes (SLNs) to determine the rate of non-SLN metastasis and axillary recurrences, and to compare actual non-SLN metastasis rates with those predicted by the Memorial Sloan-Kettering Cancer Center (MSKCC) nomogram.
We identified 116 stage I to III breast cancer patients who underwent sentinel lymph node biopsy and had micrometastases or ITCs (<2-mm deposits). Patients underwent completion axillary lymph node dissection (ALND) (group 1) or had no further axillary surgery (group 2). P < 0.05 was considered statistically significant.
Of 116 patients with micrometastases or ITCs in SLNs, 55 (47%) underwent completion ALND (group 1), and 61 (53%) had no further axillary surgery (group 2). The rate of non-SLN metastases in group 1 patients was 9 (16%) of 55, which was significantly less than that predicted by the MSKCC nomogram (median 30%, P < 0.001). Patient age, race, tumor histology, tumor grade, estrogen receptor/Her-2neu status, and lymphovascular invasion did not differ significantly between group 1 patients with positive non-SLNs and those with negative non-SLNs (P > 0.05 for each), but patients with positive non-SLNs had larger tumors (P < 0.001). No patient in group 1 experienced an axillary recurrence, while only one patient (1.6%) in group 2 experienced axillary recurrence.
The actual rate of positive non-SLNs for breast cancer patients with SLN micrometastases or ITCs who underwent completion ALND was significantly less than that predicted by the MSKCC nomogram. The rate of axillary recurrence is negligible, regardless of the extent of axillary staging.
Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that is known to be chemosensitive. In patients with TNBC, we sought to compare survival outcomes between patients receiving neoadjuvant chemotherapy, with and without complete pathologic response (pCR), and those receiving adjuvant chemotherapy.
We performed a retrospective chart review and identified 385 patients with stage I–III TNBC who were treated with neoadjuvant or adjuvant chemotherapy between 2000 and 2008. Patients were divided according to receipt of neoadjuvant chemotherapy with pCR, neoadjuvant chemotherapy without pCR, and adjuvant chemotherapy. Data were compared using Fisher’s exact test and analysis of variance (ANOVA). Kaplan–Meier curves were generated.
Of 385 patients, 151 (39%) received neoadjuvant chemotherapy and 234 (61%) received adjuvant chemotherapy. Twenty-six (17%) of those patients receiving neoadjuvant chemotherapy had pCR. After controlling for covariates associated with survival in unadjusted tests, patients undergoing neoadjuvant chemotherapy with residual tumor had significantly worse survival compared with patients receiving adjuvant therapy [hazard ratio (HR) = 0.51, P = 0.007] and a trend towards worse survival compared with patients receiving neoadjuvant therapy with pCR (HR = 0.19, P = 0.10).
Although previous clinical trials have not demonstrated a survival difference between patients receiving neoadjuvant versus adjuvant chemotherapy for breast cancer, our study suggests an overall survival benefit in patients with pCR following neoadjuvant chemotherapy compared with patients receiving adjuvant therapy. It is clear that a prospective study needs to be carried out to better elucidate the timing of chemotherapy in patients with TNBC.
The study aim was to determine the accuracy of axillary ultrasound (AUS) and fine needle aspiration biopsy (FNAB)/needle core biopsy in axillary breast cancer staging.
We reviewed 256 patients with clinically node-negative breast cancer who underwent AUS +/− FNAB/needle core biopsy. AUS-guided FNAB/needle core biopsy was compared to histopathology to determine sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV).
AUS-guided FNAB/needle core biopsy and final pathology were positive in 72/256 patients (28%). In 125/256 cases (49%), the AUS and final pathology were negative. Two of 110 patients had a false positive FNAB (1.8%); both received neoadjuvant chemotherapy. Nine patients (8%) had a false negative FNAB/needle core biopsy; the median size of lymph node metastasis was 3 mm. The sensitivity and specificity of AUS-guided FNAB/needle core biopsy was 71% and 99%, with a NPV of 84% and PPV of 97%.
AUS-guided FNAB/needle core biopsy is accurate in predicting the status of the axilla in 70% of clinically node-negative breast cancer patients. This technique is minimally invasive with a low complication rate and can obviate the need for staged lymph node procedures.
Breast cancer; Axillary ultrasound (AUS); Fine needle aspiration biopsy (FNAB)
Anti-MHC class I alloantibodies have been implicated in the process of acute and chronic rejection because these Abs can bind to endothelial cells and transduce signals leading to the activation of cell survival and proliferation pathways. To characterize the role of the MHC class I-signaling pathway in the pathogenesis of Ab-mediated rejection, we developed a mouse vascularized heterotopic cardiac allograft model in which B6.RAG1 KO hosts (H-2Kb/Db) received a fully MHC-incompatible BALB/c (H-2Kd/Dd) heart transplant and were passively transfused with anti-donor MHC class I Ab. We demonstrate that cardiac allografts of mice treated with anti-MHC class I Abs show characteristic features of Ab-mediated rejection including microvascular changes accompanied by C4d deposition. Phosphoproteomic analysis of signaling molecules involved in the MHC class I cell proliferation and survival pathways were elevated in anti-class I-treated mice compared with the isotype control-treated group. Pairwise correlations, hierarchical clustering, and multidimensional scaling algorithms were used to dissect the class I-signaling pathway in vivo. Treatment with anti-H-2Kd Ab was highly correlated with the activation of Akt and p70S6Kinase (S6K). When measuring distance as a marker of interrelatedness, multidimensional scaling analysis revealed a close association between members of the mammalian target of rapamycin pathway including mammalian target of rapamycin, S6K, and S6 ribosomal protein. These results provide the first analysis of the interrelationships between these signaling molecules in vivo that reflects our knowledge of the signaling pathway derived from in vitro experiments.
The equine MHC class I molecule was crystallized in complex with β2-microglobulin and a CTL epitope and X-ray diffraction data were collected to 2.3 Å resolution.
In order to clarify the structure and the peptide-presentation characteristics of the equine major histocompatibility complex (MHC) class I molecule, a complex of equine MHC class I molecule (ELA-A1 haplotype, 7-6 allele) with mouse β2-microglobulin and the cytotoxic T lymphocyte (CTL) epitope Env-RW12 (RVEDVTNTAEYW) derived from equine infectious anaemia virus (EIAV) envelope protein (residues 195–206) was refolded and crystallized. The crystal, which belonged to space group P21, diffracted to 2.3 Å resolution and had unit-cell parameters a = 82.5, b = 71.4, c = 99.8 Å, β = 102.9°. The crystal structure contained two molecules in the asymmetric unit. These results should help to determine the first equine MHC class I molecule structure presenting an EIAV CTL epitope.
equine MHC class I molecule
Hepatic injury due to cold storage followed by reperfusion remains a major cause of morbidity and mortality after orthotopic liver transplantation (OLT). CD4 T cell TIM-1 signaling co-stimulates a variety of immune responses in allograft recipients. This study analyzes mechanisms by which TIM-1 affects liver ischemia-reperfusion injury (IRI) in a murine model of prolonged cold storage followed by OLT. Livers from C57BL/6 mice, preserved at 4°C in UW solution for 20h, were transplanted to syngeneic recipients. There was an early (1h) increased accumulation of TIM-1+ activated CD4 T cells in the ischemic OLTs. Disruption of TIM-1 signaling with a blocking mAb (RMT1-10) ameliorated liver damage, evidenced by reduced sALT levels and well-preserved architecture. Unlike in controls, TIM-1 blockade diminished OLT expression of Tbet/IFN-γ, but amplified IL-4/IL-10/IL-22; abolished neutrophil and macrophage infiltration/activation; and inhibited NF-κB while enhancing Bcl-2/Bcl-xl. Although adoptive transfer of CD4 T cells triggered liver damage in otherwise IR-resistant RAG−/− mice, adjunctive TIM-1 blockade reduced Tbet transcription, and abolished macrophage activation, restoring homeostasis in IR-stressed livers. Further, transfer of TIM-1HiCD4+, but not TIM-1LoCD4+ T cells, recreated liver IRI in RAG−/− mice. Thus, TIM-1 expressing CD4 T cells are required in the mechanism of innate immune-mediated hepatic IRI in OLTs.
Ischemia-Reperfusion Injury; Orthotopic Liver Transplantation; TIM-1; T cell costimulation; innate immunity
Understanding human immunodeficiency virus type 1 (HIV-1) transmission is central to developing effective prevention strategies, including a vaccine. We compared phenotypic and genetic variation in HIV-1 env genes from subjects in acute/early infection and subjects with chronic infections in the context of subtype C heterosexual transmission. We found that the transmitted viruses all used CCR5 and required high levels of CD4 to infect target cells, suggesting selection for replication in T cells and not macrophages after transmission. In addition, the transmitted viruses were more likely to use a maraviroc-sensitive conformation of CCR5, perhaps identifying a feature of the target T cell. We confirmed an earlier observation that the transmitted viruses were, on average, modestly underglycosylated relative to the viruses from chronically infected subjects. This difference was most pronounced in comparing the viruses in acutely infected men to those in chronically infected women. These features of the transmitted virus point to selective pressures during the transmission event. We did not observe a consistent difference either in heterologous neutralization sensitivity or in sensitivity to soluble CD4 between the two groups, suggesting similar conformations between viruses from acute and chronic infection. However, the presence or absence of glycosylation sites had differential effects on neutralization sensitivity for different antibodies. We suggest that the occasional absence of glycosylation sites encoded in the conserved regions of env, further reduced in transmitted viruses, could expose specific surface structures on the protein as antibody targets.
We evaluated the activity and safety of the combination of topotecan, cisplatin and bevacizumab in patients with recurrent or persistent carcinoma of the cervix.
Eligible patients had persistent or recurrent cervical cancer not amenable to curative intent treatment. No prior chemotherapy for recurrence was allowed. Treatment consisted of cisplatin 50 mg/m2 day 1, topotecan 0.75 mg/m2 days 1,2 and 3 and bevacizumab 15 mg/kg day 1 every 21 days until disease progression or limiting toxicity. The primary endpoint was progression free survival at 6 months. We explored PET/CT as a potential early indicator of response to therapy.
Twenty-seven eligible patients received a median of 3 treatment cycles (range, 1–19). Median fallow-up was 10 months (range, 1.7–33.4). The 6-month PFS was 59% (80% CI: 46-70%). in 26 evaluable patients, we observed 1 CR (4%; 80% CI: 0.4–14%) and 8 PR (31%; 80% CI: 19–45%) lasting a median of 4.4 months. Ten patients had SD (39%; 80% CI: 25–53%) with median duration of 2.2 months. Median PFS was 7.1 months (80%; CI: 4.7–10.1) and median OS was 13.2 months (80% CI: 8.0–15.4). All patients were evaluated for toxicity. Grade 3–4 hematologic toxicity was common (thrombocytopenia 82% leukopenia 74%, anemia 63%, neutropenia 56%). Most patients (78%) required unanticipated hospital admissions for supportive care and/or management of toxicities.
The addition of bevacizumab to topotecan and cisplatin results in an active but highly toxic regimen. Future efforts should focus on identification of predictive biomarkers of prolonged response and regimen modifications to minimize toxicity.
Ceivical cancer; Chemotherapy; Topotecan; Cisplatin; Bevacizumab; PET/CT
Clinical trials on early stage Alzheimer’s disease (AD) are reaching a bottleneck because none of the current disease markers changes appreciably early in the disease process and therefore a huge sample is required to adequately power such trials. We propose a method to combine multiple markers so that the longitudinal rate of progression can be improved. The criterion is to maximize the probability that the combined marker will be decreased over time (assuming a negative mean slope for each marker). We propose estimates to the weights of markers in the optimum combination and a confidence interval estimate to the combined rate of progression through the maximum likelihood estimates and a bootstrap procedure. We conduct simulations to assess the performance of our estimates and compare our approach with the first principal component from a principal component analysis. The proposed method is applied to a real world sample of individuals with preclinical AD to combine measures from two cognitive domains. The combined cognitive marker is finally used to design future clinical trials on preclinical AD, demonstrating a significant improvement in reducing the sample sizes needed to power such trials when compared with individual markers alone.
Bootstrap estimate; Delta method; Multivariate random coefficients models; Power; Preclinical Alzheimer’s disease (AD); Randomized clinical trials (RCT); Sample size
There is a debate regarding the choice of operative intervention in humeral shaft fractures that require surgical intervention. The choices for operative interventions include intramedullary nailing (IMN) and dynamic compression plate (DCP). This meta-analysis was performed to compare fracture union, functional outcomes, and complication rates in patients treated with IMN or DCP for humeral shaft fractures and to develop GRADE (Grading of Recommendations, Assessment, Development, and Evaluation)-based recommendations for using the procedures to treat humeral shaft fractures. A systematic search of all the studies published through December 2012 was conducted using the Medline, Embase, Sciencedirect, OVID and Cochrane Central databases. The randomized controlled trials (RCTs) and quasi-RCTs that compared IMN with DCP in treating adult patients with humeral shaft fractures and provided data regarding the safety and clinical effects were identified. The demographic characteristics, adverse events and clinical outcomes were manually extracted from all of the selected studies. Ten studies that included a total of 448 patients met the inclusion criteria. The results of a meta-analysis indicated that both IMN and DCP can achieve similar fracture union with a similar incidence of radial nerve injury and infection. IMN was associated with an increased risk of shoulder impingement, more restriction of shoulder movement, an increased risk of intraoperative fracture comminution, a higher incidence of implant failure, and an increased risk of re-operation. The overall GRADE system evidence quality was very low, which reduces our confidence in the recommendations of this system. DCP may be superior to IMN in the treatment of humeral shaft fractures. Because of the low quality evidence currently available, high-quality RCTs are required.
AIM: To evaluate the impact of Bmi-1 on cell senescence and metastasis of human gastric cancer cell line BGC823.
METHODS: Two pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the Bmi-1 gene were designed, synthesized, annealed and cloned into the pRNAT-U6.2 vector. After DNA sequencing to verify the correct insertion of the shRNA sequences, the recombinant plasmids were transfected into BGC823 cells. The expression of Bmi-1 mRNA and protein was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The effects of Bmi-1 knockdown on cell senescence and metastasis were determined by the β-Gal activity assay and Boyden chamber assay, respectively.
RESULTS: The double-stranded oligonucleotide fragments of Bmi-1 short interfering RNA (siRNA) cloned into pRNAT-U6.2 vector conformed to the inserted sequence. RT-PCR and Western blotting indicated that the expression levels of Bmi-1 gene mRNA and protein were markedly decreased in transfected BGC823 cells with pRNAT-U6.2-si1104 and pRNAT-U6.2-si1356, especially in transfected BGC823 cells with pRNAT-U6.2-si1104, compared with two control groups (empty vector and blank group). In particular, Bmi-1 protein expression was almost completely abolished in cells transfected with the recombinant vector harboring shRNA targeting the sequence GGAGGAGGTGAATGATAAA (nt1104-1122). Compared with untransfected cells and cells transfected with the empty vector, the mean percentage of senescent cells increased and the number of cells passing through the Matrigel decreased in cells transfected with the recombinant vectors.
CONCLUSION: Silencing Bmi-1 by RNA interference can increase the senescent cell rate and effectively reduce the metastasis of gastric cancer cells.
Bmi-1; Gastric cancer; Senescence; Metastasis
Lymphotoxin-alpha (LTA) is a pro-inflammatory cytokine that plays an important role in the inflammatory and immunologic response. Numerous studies have shown LTA polymorphisms as risk factors for cancers, but the results remain inconclusive. The goal of the present meta-analyses is to establish the associations between cancers and four LTA variants (rs1041981, rs2239704, rs2229094 and rs746868). A total of 30 case-control studies involving 58,649 participants were included in the current meta-analyses. Our results showed significant associations with increased cancer risk for rs1041981 (odd ratio (OR) = 1.15, 99% confidential interval (CI) = 1.07-1.25, P < 0.0001, I2 = 12.2%), rs2239704 (OR = 1.08, 99% CI = 1.01-1.16, P = 0.021, I2 = 0.0%) and rs2229094 (OR = 1.28, 99% CI = 1.09-1.50, P = 0.003, I2 = 0.0%). No evidence was found for the association between rs746868 and cancer risk (OR = 1.01, 99% CI = 0.93-1.10, P = 0.771, I2 = 0.0%). Subgroup meta-analysis suggested that rs2239704 was likely to increase the risk of hematological malignancy (OR = 1.10, 99% CI = 1.01–1.20, P = 0.023, I2 = 0.0%), and rs2229094 was specific for the increased risk of adenocarcinoma (OR = 1.33, 99% CI = 1.11-1.59, P = 0.002, I2 = 0.0%). In conclusion, our meta-analyses suggested that the LTA rs1041981, rs2239704 and rs2229094 polymorphisms contributed to the increased risk of cancers. Future functional studies were needed to clarify the mechanistic roles of the three variants in the cancer risk.
The identification of mammalian target of rapamycin (mTOR) as a major mediator of neurofibromatosis-1 (NF1) tumor growth has led to the initiation of clinical trials using rapamycin analogs. Previous studies from our laboratory have shown that durable responses to rapamycin treatment in a genetically-engineered mouse (GEM) model of Nf1 optic glioma require 20mg/kg/day, whereas only transient tumor growth suppression was observed with 5mg/kg/day rapamycin despite complete silencing of ribosomal S6 activity. To gain clinically-relevant insights into the mechanism underlying this dose-dependent effect, we employed Nf1-deficient glial cells in vitro and in vivo. First, there is an exponential relationship between blood and brain rapamycin levels. Second, we demonstrate that currently-used biomarkers of mTOR pathway inhibition (phospho-S6, phospho-4EBP1, phospho-STAT3, and Jagged-1 levels) and tumor proliferation (Ki67) do not accurately reflect mTOR target inhibition or Nf1-deficient glial growth suppression. Third, the incomplete suppression of Nf1-deficient glial cell proliferation in vivo following 5mg/kg/day rapamycin treatment reflects mTOR-mediated AKT activation, such that combined 5mg/kg/day rapamycin and PI3-Kinase (PI3K) inhibition or dual PI3K/mTOR inhibition recapitulates the growth suppressive effects of 20mg/kg/day rapamycin. These new findings argue for the identification of more accurate biomarkers for rapamycin treatment response, and provide reference preclinical data for comparing human rapamycin levels with target effects in the brain.
glioma; astrocytoma; S6; preclinical; neurofibromin
Following mucosal human immunodeficiency virus type 1 (HIV-1) transmission, type 1 interferons (IFNs) are rapidly induced at sites of initial virus replication in the mucosa and draining lymph nodes. However, the role played by IFN-stimulated antiviral activity in restricting HIV-1 replication during the initial stages of infection is not clear. We hypothesized that if type 1 IFNs exert selective pressure on HIV-1 replication in the earliest stages of infection, the founder viruses that succeed in establishing systemic infection would be more IFN-resistant than viruses replicating during chronic infection, when type 1 IFNs are produced at much lower levels. To address this hypothesis, the relative resistance of virus isolates derived from HIV-1-infected individuals during acute and chronic infection to control by type 1 IFNs was analysed.
The replication of plasma virus isolates generated from subjects acutely infected with HIV-1 and molecularly cloned founder HIV-1 strains could be reduced but not fully suppressed by type 1 IFNs in vitro. The mean IC50 value for IFNα2 (22 U/ml) was lower than that for IFNβ (346 U/ml), although at maximally-inhibitory concentrations both IFN subtypes inhibited virus replication to similar extents. Individual virus isolates exhibited differential susceptibility to inhibition by IFNα2 and IFNβ, likely reflecting variation in resistance to differentially up-regulated IFN-stimulated genes. Virus isolates from subjects acutely infected with HIV-1 were significantly more resistant to in vitro control by IFNα than virus isolates generated from the same individuals during chronic, asymptomatic infection. Viral IFN resistance declined rapidly after the acute phase of infection: in five subjects, viruses derived from six-month consensus molecular clones were significantly more sensitive to the antiviral effects of IFNs than the corresponding founder viruses.
The establishment of systemic HIV-1 infection by relatively IFNα-resistant founder viruses lends strong support to the hypothesis that IFNα plays an important role in the control of HIV-1 replication during the earliest stages of infection, prior to systemic viral spread. These findings suggest that it may be possible to harness the antiviral activity of type 1 IFNs in prophylactic and potentially also therapeutic strategies to combat HIV-1 infection.
Human immunodeficiency virus type 1; Type 1 interferon; Viral inhibition; Founder virus; Acute infection