[Arterial prophylactic occlusion technique in the application of surgery for locally advanced pancreatic cancer with arterial involvement after conversion therapy].

Objective: To investigate and compare the clinical outcomes of the arterial pre-occlusion technique(APOT) and the traditional technique in the surgery of locally advanced pancreatic cancer with arterial involvement after conversion therapy. Methods: This is a retrospective cohort study. The clinical data of 145 patients with locally advanced pancreatic cancer with arterial involvement admitted to the Department of Hepato-Biliary-Pancreatic Surgery of the First Hospital Affiliated to Naval Medical University,from January 2020 to December 2022 were retrospectively analyzed. All patients completed neoadjuvant therapy for tumors, and the feasibility of radical surgical treatment was determined by a multidisciplinary collaborative team evaluation before surgery. According to whether the intraoperative artery was pre-occluded, 145 patients were divided into two groups, including 28 cases in the APOT group(16 males, 12 females, aged (59.0±9.4) years), and 117 cases in the routine surgery group(76 males, 41 females, aged (55.1±8.2) years). To ensure comparability of baseline data between the APOT group and the routine surgery group, a 1∶2 match was performed using the propensity score matching method, and the caliper value was 0.006 45. The t-test,the Mann-Whitney U test, χ2 test or Fisher's exact test were used to compare the data between the two groups,respectively. Results: After matching the propensity score,there were 28 cases in the APOT group and 56 cases in the routine surgery group. There were no significant differences in gender,age,preoperative comorbidities,preoperative body mass index,surgical approaches,chemotherapy regimen,stereotactic body radiation therapy ratio,tumor markers,and type of invaded artery between the two groups (all P>0.05).The arterial occlusion time M(IQR) in the APOT group was 7.0(3.8)minutes(range:3 to 15 minutes),and no ischemic manifestations were observed in the distal target organs that blocked blood vessels after surgery. The operation time was (170.3±57.7)minutes in the APOT group and (235.0±80.2)minutes in the routine surgery group,and the difference was statistically significant (t=-3.800,P<0.01). The APOT group also experienced less intraoperative blood loss(650(588)ml vs. 800(600)ml;U=1 026.500,P=0.021). No significant differences were found between the groups in combined vein resection and reconstruction,celiac trunk resection,early postoperative complications, readmission rates at 30 days,and postoperative length of stay(all P>0.05). Extra-arterial dissection was performed in all patients,with arterial resection and reconstruction in 3 cases: 2 cases in the APOT group(1 case involving the superior mesenteric artery and 1 case involving the common hepatic artery) and 1 case in the routine group(involving the common hepatic artery). Postoperative abdominal bleeding occurred in 4 cases,with 3 cases in the routine group,1 case in the routine group. The R0 resection rate was 85.7%(24/28) in the APOT group and 80.4%(45/56) in the routine group,without significant differences between the groups(P=0.763). The median overall survival time was 27.6 months for the APOT group and 22.5 months for the routine group,while the median disease-free survival was 11.7 months and 16.8 months,respectively,with no significant differences between the two groups(P=0.532,P=0.927). Conclusion: The arterial pre-occlusion technique can be used for extra-arterial dissection in patients with locally advanced pancreatic cancer involving the arteries,reducing surgery time and intraoperative blood loss.

Clinical application of flash spiral mode of high-pitch dual source CT in carotid, cardiac and cerebral vessels combined one-stop angiography.

OBJECTIVE: This study aims to evaluate the clinical application value of flash spiral mode of high-pitch dual source CT in carotid, cardiac and cerebral vessels combined one-stop imaging. PATIENTS AND METHODS: A total of 100 consecutive patients were given carotid, cardiac and cerebral vessels combined one-stop imaging at flash spiral mode of high-pitch dual source CT. 27 patients received DSA examination of carotid and cerebral vessels, and 38 patients received digital subtraction angiography (DSA) examination of the coronary artery at the same time. Carotid, cardiac and cerebral vessels combined one-stop imaging was compared with "golden standard", DSA image. RESULTS: The overall satisfaction rate of coronary arteries, extracranial segment of the carotid artery (CA-E), intracranial segment of the carotid artery (CA-I), and cerebral vessels (anterior, middle, and posterior cerebral artery) were 93%, 99%, 95% and 97% respectively, and the positive rate of hemadostenosis was consistent with DSA. The kappa value indicating consistency of cerebral, carotid and coronary artery vessels was 0.78382, 0.80654, 0.82398, respectively. CONCLUSIONS: The method of carotid, cardiac and cerebral vessels combined one-stop imaging by flash spiral mode of high-pitch dual source CT can provide high image quality. It comprehensively evaluates stenosis of carotid, cardiac and cerebral vessels, which is of great importance for early intervention in harmful events of the cardia and cerebrovascular disorder.

Few-Body Perspective of a Quantum Anomaly in Two-Dimensional Fermi Gases.

A quantum anomaly manifests itself in the deviation of the breathing mode frequency from the scale invariant value of 2ω in two-dimensional harmonically trapped Fermi gases, where ω is the trapping frequency. Its recent experimental observation with cold atoms reveals an unexpected role played by the effective range of interactions, which requires a quantitative theoretical understanding. Here we provide accurate, benchmark results on a quantum anomaly from a few-body perspective. We consider the breathing mode of a few trapped interacting fermions in two dimensions up to six particles and present the mode frequency as a function of scattering length for a wide range of effective range. We show that the maximum quantum anomaly gradually reduces as the effective range increases while the maximum position shifts towards the weak-coupling limit. We extrapolate our few-body results to the many-body limit and find a good agreement with the experimental measurements. Our results may also be directly applicable to a few-fermion system prepared in microtraps and optical tweezers.

Cluster Formation in Two-Component Fermi Gases.

Two-component fermions are known to behave like a gas of molecules in the limit of Bose-Einstein condensation of diatomic pairs tightly bound with zero-range interactions. We discover that the formation of cluster states occurs when the effective range of two-body interaction exceeds roughly 0.46 times the scattering length, regardless of the details of the short-range interaction. Using an explicitly correlated Gaussian basis set expansion approach, we calculate the binding energy of cluster states in trapped few-body systems and show the difference of structural properties between cluster states and gaslike states. We identify the condition for cluster formation and discuss the potential observation of cluster states in experiments.

[Aspirin inhibits cell stemness of esophageal cancer by downregulation of chemokine CCL2].

Objective: To investigate the effect of tumor-associated macrophages on the stemness of esophageal cancer cells and the potential mechanism of antiproliferative effects of aspirin (ASA). Methods: The effects of aspirin on the stemness characteristics of KYSE-450 cells and KYSE-450 cells co-cultured with M2 macrophages (KYSE-450+ M2) were performed using spheroid formation assay. After treatment with aspirin, the expression of different chemokines, the core pluripotency gene Nanog and the stem cell marker CD90 in different cell groups were determined by real-time quantitative PCR, flow cytometry and Western blot. Results: The number of spheres formed in the ASA and KYSE-450+ M2 cell groups were 7.00±1.23 and 34.33±2.33, respectively, showing statistically significant difference compared with that of control group (14.50±2.33, all P<0.05). The number of spheres in KYSE-450+ M2+ ASA cell group were 20.67±2.33, which was significantly lower than that of KYSE-450+ M2 group (P<0.05). The expression levels of Nanog gene in control and ASA groups were 1.00 and 0.50±0.10, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression of Nanog gene in cells of KYSE-450+ M2 group and M2+ KYSE-450+ ASA group was 1.74±0.13 and 1.43±0.05, showing statistically significant difference (P<0.05). When chemokine CCL2 was knocked down, the levels of Nanog gene in M2+ shCCL2-KYSE450+ ASA group and M2+ shCCL2-KYSE450 group were decreased to 1.22±0.11 and 1.17±0.08, respectively, and there was no statistically significant difference between them (P=0.69). Flow cytometry analyses showed that the expression levels of CD90 in control and ASA cells were (2.93±0.52)% and (1.30±0.17)%, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression levels of CD90 in M2+ shCCL2-KYSE450 cells and M2+ shCCL2-KYSE450+ ASA cells were (4.07±0.12)% and (4.73±0.38)%, respectively, showing no statistically significant difference (P=0.17). Conclusions: Tumor-associated macrophages enhances the stemness of esophageal cancer cells, whereas aspirin attenuates the stemness by suppressing the expression of CCL2. Aspirin plays an anti-tumor effect in esophageal cancer cells.

Expression of three proteins in endometrioid adenocarcinoma and their significance in clinical nursing, diagnosis and treatment.

OBJECTIVE: This study aimed to investigate the role of NEDD9 (neural precursor cell expressed developmentally down-regulated 9), BCAR1/P130CAS (BCAR1/P130 Crk-associated substrate) and paxillin in predicting the prognosis of endometrioid adenocarcinoma (EA), so as to guild the nursing of EA. PATIENTS AND METHODS: A total of 65 patients who visited Affiliated Hongqi Hospital of Mudanjiang Medical University between June 2015 and June 2017 were enrolled. They underwent gynecological surgery and had their EA confirmed by pathology, and they were assigned to the EA group. All EA tissues were sampled and archived in paraffin blocks. In addition, 40 specimens of atypical endometrial hyperplasia (EAH) (the EAH group) and 40 specimens of normal proliferative endometria with benign uterine fibroids (the EN group) were selected as controls. The protein levels of NEDD9, BCAR1/P130CAS, and paxillin in each group were then detected by immunohistochemical staining. RESULTS: The expression of the three proteins in the EA group and EAH group was significantly higher than that in the EN group, and their expression was significantly correlated with the clinical stage, histological grade and lymph node metastasis of EA. In addition, the expression of NEDD9, BCAR1/P130CAS, and paxillin in the EA group was positively correlated with each other. CONCLUSIONS: BCAR1/P130CAS and paxillin interact with NEDD9 to participate in the growth and migration of EA cells. Therefore, their proteins can be used as biomarkers for the diagnosis, treatment, and prognosis of EA.

Laboratory studies of ultrasonic wave response of fractures with different lengths: Anisotropy characteristics and coda analysis.

Ultrasonic wave propagation in a synthetic medium with different scale fractures is investigated through the physics modeling research. The experiment model consisting of two layers, the first layer is divided into four fractured blocks with a constant fracture density but different dimension and a block without fractures as the reference region. The velocity and reflected amplitude of wave derived from the wide azimuth data processing are anisotropic due to the fractured layer. Coda wave characteristics versus offset and azimuth are obviously different for varying fracture scales. The ratio of wavelength and fracture dimension is an important parameter in the wave-fracture interaction based on the multiple scattering theory. When the wavelength of the incident wave is close to the fracture length, the scattering is dominant in the shot records and the waves are slowed and attenuated largely for the reflections from the bottoms of fractured layer and base model. The physical modeling results demonstrate that it is possible to extract fracture dimension and orientation information by analyzing code and primary waves from the reflection data.

Single cell transcriptome profiling revealed differences in gene expression during oocyte maturation in Haimen white goats.

Juvenile in vitro embryo transfer is an important animal reproductive technology that can shorten the generation interval of livestock, explore the reproductive potential of dams with excellent genetic traits, accelerate genetic progress and production efficiency of the herd, and provide a wealth of genetic resources for livestock breeding. However, oocytes from kids do not develop as well as those from female goats during in vitro maturation. To identify differences during different stages of oocyte maturation, we used single cell transcriptome sequencing to compare gene expression in mature oocytes from kids and female goats. We identified 1086 differentially expressed genes in mature oocytes from kids and female goats. Of these, we observed upregulated expression in 355 genes and downregulated expression in 435 genes. The differentially expressed genes were involved in a total of 245 different pathways; of which 30 were significant (P ≤ 0.05). We used real-time quantitative polymerase chain reaction to screen and verify the expression of five genes specifically involved in oocyte maturation (MOS, RPS6KA1, CPEB1, ANAPC13, and CDK1). Further study of these genes will be of great importance for improving the reproductive performance of Haimen white goats.

Multiple variants in 5q31.1 are associated with systemic lupus erythematosus susceptibility and subphenotypes in the Han Chinese population.

BACKGROUND: A previous study provided evidence for a genetic association between PPP2CA on 5q31.1 and systemic lupus erythematosus (SLE) across multi-ancestral cohorts, but failed to find significant evidence for an association in the Han Chinese population. OBJECTIVES: To explore the association between this locus and SLE using data from our previously published genome-wide association study (GWAS). METHODS: Single-nucleotide polymorphisms (SNPs) rs7726414 and rs244689 (near TCF7 and PPP2CA in 5q31.1) were selected as candidate independent associations from a large-scale study in a Han Chinese population consisting of 1047 cases and 1205 controls. Subsequently, 3509 cases and 8246 controls were genotyped in two further replication studies. We then investigated the SNPs' associations with SLE subphenotypes and gene expression in peripheral blood mononuclear cells. RESULTS: Highly significant associations with SLE in the Han Chinese population were detected for SNPs rs7726414 and rs244689 by combining the genotype data from our previous GWAS and two independent replication cohorts. Further conditional analyses indicated that these two SNPs contribute to disease susceptibility independently. A significant association with SLE, age at diagnosis < 20 years, was found for rs7726414 (P = 0·001). The expression levels of TCF7 and PPP2CA messenger RNA in patients with SLE were significantly decreased compared with those in healthy controls. CONCLUSIONS: This study found evidence for multiple associations with SLE in 5q31.1 at genome-wide levels of significance for the first time in a Han Chinese population, in a combined genotype dataset. These findings suggest that variants in the 5q31.1 locus not only provide novel insights into the genetic architecture of SLE, but also contribute to the complex subphenotypes of SLE.

Suppression of collagen-induced arthritis by lipopolysaccharide in DBA/1 mice.

OBJECTIVE: Injection of lipopolysaccharide (LPS) has both promotion and inhibition effects on the autoimmune disease. Given the variable roles of LPS in autoimmune diseases, the role of LPS played in collagen-induced arthritis (CIA, autoimmune disease) model remains to be further determined. MATERIALS AND METHODS: CIA was induced by intradermal injection of collagen type II (CII) in DBA/1 mice (day 0) followed by a booster injection on day 21. Mice of CIA with LPS injection group (CIA+ LPS group) were intraperitoneally injected with 50 µg LPS on day 42. Tissues such as carpal joints and fingers were stained with hematoxylin and eosin (H&E) for histopathology analysis. Inflammation, pannus formation and bone resorption were monitored by a macroscopic scoring system. Serum level of IgG2a antibody was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The incidence of arthritis in CIA group was much higher than that in CIA+ LPS group (100%: 46.5%, p < 0.05), as same as the arthritis score (5.38:1.37, p = 8.16 × 10-6). Besides, the histopathologic score was also higher in CIA group than that in CIA+ LPS group (15.0:5.36). Compared with CIA group, mild synovial hyperplasia and no articular cartilage damage were observed in CIA+ LPS group. Besides, mice of CIA group produced a significantly higher level of IgG2a than CIA+ LPS group (3922 ng/ml: 2084 ng/ml, p = 0.0333) when arthritis developed. CONCLUSIONS: Our findings showed that LPS might suppress CIA progression under special conditions, opening up a new understanding of the roles of LPS in arthritis and new possibilities for a clinical therapy of CIA.

Genetic diversity and taxonomic status of Gymnocypris chilianensis based on the mitochondrial DNA cytochrome b gene.

In order to study the genetic diversity and taxonomic status of Gymnocypris chilianensis on a molecular level, the mitochondrial DNA cytochrome b gene was sequenced for 74 individuals of G. chilianensis from two locations (Heihe River and Shule River) and 42 individuals of its affinis species Gymnocypris przewalskii. Analyses of genetic diversity and sequence differences were conducted for these samples, combined with the analysis of 30 homologous sequences of another affinis species Gymnocypris eckloni, which were downloaded from GenBank. The results showed that both the haplotype diversity (h = 0.9820) and nucleotide diversity (π= 0.0039) of the Shule River G. chilianensis were lower than the other populations, thus, the Shule River G. chilianensis should be prioritized for protection because of its lower genetic diversity level. The results of sequence analysis showed that the genetic distance between the Heihe River G. chilianensis population and the Shule River G. chilianensis population was 0.0064, and the genetic distance between these two populations and the G. przewalskii population was 0.0838 and 0.0810, respectively. The genetic distance between the two G. chilianensis populations and the G. eckloni population was 0.0805 and 0.0778, respectively. Analysis of sequence differences indicates that G. chilianensis is sufficiently diverged from G. przewalskii and G. eckloni to the extent that it has reached species level, thus, G. chilianensis can be considered an independent species of Gymnocypris.

HTRA3 is reduced in ovarian cancers regardless of stage.

Ovarian cancer is the leading cause of death in gynaecological cancers. The high temperature requirement factor A3 (HtrA3) is involved in the pathogenesis of ovarian cancer. In this study we investigated whetherHtrA3 protein levels were altered in subtypes of ovarian cancer and whether HtrA3 down-regulation was associated with peritoneal metastasis. Ovarian cancer tissues from 89 patients were analyzed by immunohistochemistry. The levels of HtrA3 protein were lower in all subtypes of ovarian cancer and the lowest levels of HtrA3 were in epithelial ovarian cancer. The down-regulation of HtrA3 levels was not correlated with peritoneal metastasis of epithelial ovarian cancer.

Extratemporal abnormalities of brain parenchyma in young adults with temporal lobe epilepsy: a diffusion tensor imaging study.

AIM: To examine extratemporal abnormalities of the cerebral parenchyma in young adult temporal lobe epilepsy (TLE) patients using diffusion tensor imaging (DTI). MATERIALS AND METHODS: The study comprised 20 adults with unilateral TLE and 20 controls. The fractional anisotropy (FA), apparent diffusion coefficient (ADC), parallel eigenvalue (λ∥), and perpendicular eigenvalue (λ⊥) were calculated in the regions of interest (ROIs) using a 3 T MRI system. ROIs included the anterior/posterior limb of the internal capsule (AIC/PIC), external capsule (EC), head of caudate nucleus (HCN), lenticular nucleus (LN), thalamus (TL), and genu/body/splenium of the corpus callosum (GCC/BCC/SCC). RESULTS: Compared to controls, TLE patients showed lower FA in all ROIs; higher ADC in bilateral ECs, HCNs, TLs, and BCC; lower λ∥ in the ipsilateral LN and bilateral AICs, TL, and GCC; and higher λ⊥ in all ROIs except the bilateral PICs. In TLE patients, the ipsilateral TL had decreased FA compared with the contralateral TL. Pearson correlation analysis revealed a negative correlation between the ADC of the GCC and the age at onset of epilepsy; the λ∥ of the ipsilateral PIC and age at onset of epilepsy; the λ⊥ of the contralateral AIC and duration of epilepsy, respectively; and a positive correlation between the ADC of the GCC and the duration of epilepsy and the λ⊥ of the GCC and the duration of epilepsy, respectively. CONCLUSION: The study revealed bilateral extratemporal abnormalities in young adult TLE patients compared with controls. In addition, TLE patients with younger age at onset or longer duration of epilepsy may have more serious extratemporal changes.

Prognostic impact of pERK in advanced hepatocellular carcinoma patients treated with sorafenib.

BACKGROUND: Sorafenib represents the standard of care targeted therapy for patients with advanced hepatocellular carcinoma (HCC). However, biomolecules that predict a patient's response to sorafenib treatment for HCC remain largely unknown. Thus, this study was designed to investigate whether phosphorylated ERK (pERK) and members of the sorafenib target or PI3K/Akt/mTOR signaling pathway predict the efficacy of sorafenib in advanced HCC patients. METHODOLOGY: From December 2008 to October 2011, pathological specimens from 54 advanced HCC patients received sorafenib treatment were obtained. Clinicopathological variables, treatment response, survival and time to progression (TTP) were recorded. Immunophenotypical analysis was carried out using antibodies against pERK, phosphorylated S6K (pS6K), VEGFR2 and PTEN. RESULTS: The median overall survival (OS) and TTP were 14.2 and 3.4 months, respectively, and the disease control rate (DCR) was 59.3%. Better Eastern Cooperative Oncology Group Performance Status (ECOG PS) (95% CI: 3.27-4.93 m vs. 1.15-2.85 m, p = 0.01), Child-Pugh class A score (95% CI: 3.47-4.53 vs. 1.14-2.06 m, p < 0.01), and higher pERK (3.34-6.66 m vs. 1.33-2.67 m, p = 0.03) and VEGFR2 (3.49-6.52 m vs. 2.15-2.73 m, p = 0.04) immunohistochemical staining score were associated with increased TTP by univariate analysis. The ECOG PS (p = 0.022), Child-Pugh class (p = 0.045) and pERK staining score (p = 0.012) were found to be associated with TTP using multivariate analysis. CONCLUSION: Sorafenib treatment outcome is favorable in advanced HCC patients who received tumor resection and who have a good ECOG PS and Child-Pugh class A liver function. The pERK immunohistological staining score, ECOG PS and Child-Pugh class may be helpful in determining patients most likely to benefit from sorafenib therapy.

Gene-gene interactions in IL23/Th17 pathway contribute to psoriasis susceptibility in Chinese Han population.

BACKGROUND: Psoriasis is a common chronic inflammatory skin disease. IL23/Th17 is a newly confirmed pathway in psoriasis. OBJECTIVE: To investigate the gene-gene interactions in IL23/Th17 pathway underlying psoriasis. METHODS: A total of 299 single-nucleotide polymorphisms from 11 genes in IL23/Th17 pathway were genotyped on 1139 patients with psoriasis and 1694 controls. Multifactor dimensionality reduction and logistic regression algorithms were applied to explore the gene-gene interactions. RESULTS: We found that there were a three-way interaction among IL21, CCR4 and TNF(χ(2) = 5.02(1), P = 0.025) and three pair-wise gene-gene interactions between IL12RB1 and CCR4(χ(2) = 11.66(4), P = 0.0201), IL22 and CCR4 (χ(2) = 11.97(4), P = 0.0176), IL12RB1 and IL6 (χ(2) = 7.31(1), P = 0.0069) in psoriasis. CONCLUSIONS: Our results might be helpful for explaining the missing heritability of the psoriasis due to epistasis and provide a deep insight into the important role of the IL23/Th17 pathway in the pathogenesis of psoriasis.

A single-nucleotide polymorphism rs4639966 in 11q23.3 is associated with clinical features of systemic lupus erythematosus in the Chinese population.

In our previous genome-wide association study (GWAS), we identified an association signal of the single-nucleotide polymorphism (SNP) rs4639966 (p = 1.25 × 10(-16), odds ratio [OR] = 1.29) within 11q23.3. The aim of this study was to investigate its relationship with disease subphenotypes, including renal nephritis, photosensitivity, antinuclear antibody (ANA), age at diagnosis, malar rash, discoid rash, immunological disorder, oral ulcer, hematological disorder, neurological disorder, serositis, arthritis and vasculitis. In this study, we used 4199 cases and 8255 controls from our previous GWAS to explore the association between 11q23.3 with subphenotypes of systemic lupus erythematosus (SLE). Data were analyzed with PLINK 1.07 software. Significant associations were found for the SNP rs4639966 of 11q23.3 with SLE of age at diagnosis <20 years (OR = 1.18, p = 0.0049), malar rash (OR = 1.13, p = 0.01) and vasculitis (OR = 1.17, p = 0.02). The study suggested that 11q23.3 might not only play important roles in the development of SLE, but also contribute to the complex phenotypes of SLE.

Different cellular p16(INK4a) localisation may signal different survival outcomes in head and neck cancer.

BACKGROUND: Recently, the management of head and neck squamous cell carcinoma (HNSCC) has focused considerable attention on biomarkers, which may influence outcomes. Tests for human papilloma infection, including direct assessment of the virus as well as an associated tumour suppressor gene p16, are considered reproducible. Tumours from familial melanoma syndromes have suggested that nuclear localisation of p16 might have a further role in risk stratification. We hypothesised p16 staining that considered nuclear localisation might be informative for predicting outcomes in a broader set of HNSCC tumours not limited to the oropharynx, human papilloma virus (HPV) status or by smoking status. METHODS: Patients treated for HNSCC from 2002 to 2006 at UNC (University of North Carolina at Chapel Hill) hospitals that had banked tissue available were eligible for this study. Tissue microarrays (TMA) were generated in triplicate. Immunohistochemical (IHC) staining for p16 was performed and scored separately for nuclear and cytoplasmic staining. Human papilloma virus staining was also carried out using monoclonal antibody E6H4. p16 expression, HPV status and other clinical features were correlated with progression-free (PFS) and overall survival (OS). RESULTS: A total of 135 patients had sufficient sample for this analysis. Median age at diagnosis was 57 years (range 20-82), with 68.9% males, 8.9% never smokers and 32.6% never drinkers. Three-year OS rate and PFS rate was 63.0% and 54.1%, respectively. Based on the p16 staining score, patients were divided into three groups: high nuclear, high cytoplasmic staining group (HN), low nuclear, low cytoplasmic staining group (LS) and high cytoplasmic, low nuclear staining group (HC). The HN and the LS groups had significantly better OS than the HC group with hazard ratios of 0.10 and 0.37, respectively, after controlling for other factors, including HPV status. These two groups also had significantly better PFS than the HC staining group. This finding was consistent for sites outside the oropharynx and did not require adjustment for smoking status. CONCLUSION: Different p16 protein localisation suggested different survival outcomes in a manner that does not require limiting the biomarker to the oropharynx and does not require assessment of smoking status.