Identification of characteristic aroma compounds in chicken meat and their metabolic mechanisms using gas chromatography-olfactometry, odor activity values, and metabolomics.

Aroma has an important influence on the aroma quality of chicken meat. This study aimed to identify the characteristic aroma substances in chicken meat and elucidate their metabolic mechanisms. Using gas chromatography-olfactometry and odor activity values, we identified nonanal, octanal, and dimethyl tetrasulfide as the basic characteristic aroma compounds in chicken meat, present in several breeds. Hexanal, 1-octen-3-ol, (E)-2-nonenal, heptanal, and (E,E)-2,4-decadienal were breed-specific aroma compounds found in native Chinese chickens but not in the meat of white-feathered broilers. Metabolomics analysis showed that L-glutamine was an important metabolic marker of nonanal, hexanal, heptanal, octanal, and 1-octen-3-ol. Exogenous supplementation experiments found that L-glutamine increased the content of D-glucosamine-6-P and induced the degradation of L-proline, L-arginine, and L-lysine to enhance the Maillard reaction and promote the formation of nonanal, hexanal, heptanal, octanal, and 1-octen-3-ol, thus improving the aroma profile of chicken meat.

Quality assurance in a phase III, multicenter, randomized trial of POstmastectomy radioThErapy in Node posiTive breast cancer with or without Internal mAmmary nodaL irradiation (POTENTIAL): a planning benchmark case.

PURPOSE: To report the planning benchmark case results of the POTENTIAL trial-a multicenter, randomized, phase 3 trial-to evaluate the value of internal mammary nodal (IMN) irradiation for patients with high-risk breast cancer. METHODS: All participating institutions were provided the outlines of one benchmark case, and they generated radiation therapy plans per protocol. The plans were evaluated by a quality assurance team, after which the institutions resubmitted their revised plans. The information on beams arrangement, skin flash, inhomogeneity corrections, and protocol compliance was assessed in the first and final submission. RESULTS: The plans from 26 institutions were analyzed. Some major deviations were found in the first submission. The protocol compliance rates of dose coverage for the planning target volume of chest wall, supraclavicular fossa plus axilla, and IMN region (PTVim) were all significantly improved in the final submission, which were 96.2% vs. 69.2%, 100% vs. 76.9%, and 88.4% vs. 53.8%, respectively. For OARs, the compliance rates of heart Dmean, left anterior descending coronary artery V40Gy, ipsilateral lung V5Gy, and stomach V5Gy were significantly improved. In the first and final submission, the mean values of PTVim V100% were 79.9% vs. 92.7%; the mean values of heart Dmean were 11.5 Gy vs. 9.7 Gy for hypofractionated radiation therapy and 11.5 Gy vs. 11.0 Gy for conventional fractionated radiation therapy, respectively. CONCLUSION: The major deviations were corrected and protocol compliance was significantly improved after revision, which highlighted the importance of planning benchmark case to guarantee the planning quality for multicenter trials.

High sensitivity gas sensor based on surface exciton polariton enhanced photonic spin Hall effect.

In this paper, the sub-wavelength transverse displacement of photonic spin Hall effect (PSHE) is significantly enhanced by the surface exciton polariton (SEP) for application in gas sensing. The transverse displacement of 14.4 times the wavelength of incident light is achieved with the SEP enhanced PSHE, which is about 3 times that of surface plasmon resonance enhanced PSHE. A gas sensor based on SEP enhanced PSHE is proposed for the detection of SO2, and the refractive index sensitivity of 6320.4 µm/RIU is obtained in the refractive index range from 1.00027281 to 1.00095981. These results undoubtedly demonstrate SEP to be a promising mechanism for PSHE enhancement, and open up new opportunities for highly sensitive gas sensing, biosensing, and chemical sensing.

Variability of Target Volumes and Organs at Risk Delineation in Breast Cancer Radiation Therapy: Quality Assurance Results of the Pretrial Benchmark Case for the POTENTIAL Trial.

PURPOSE: To estimate the variations in clinical target volumes (CTVs) and organs at risk delineation within the quality assurance (QA) program of the POTENTIAL trial, which is a multicenter, randomized phase 3 trial evaluating postmastectomy radiation therapy (RT), with or without internal mammary nodal irradiation, for patients with high-risk breast cancer. METHODS AND MATERIALS: The simulating computed tomography scan data set of a benchmark case was sent to the participating centers, and the delineation of CTVs and organs at risk was required to be completed by the investigators following protocol guidelines. All submitted contours were reviewed and compared with the reference contours created by the QA team, using quantitative geometric analysis regarding volume and the Jaccard Index (JCI), Dice similarity coefficient, Geographic Miss Index, Discordance Index, and mean distance to agreement. In addition to the whole-volume analysis of all structures, the combination contour of the supraclavicular fossa and level III and II axilla (CTVsc + axIII + axII) was further analyzed on a slice-by-slice basis. RESULTS: The contours from 26 centers were reviewed and variations were observed between submission and reference. The variations of the CTV of the chest wall, contralateral breast, and heart were small, for which the mean JCI values were 0.62, 0.68, and 0.87, respectively. However, the mean JCI values of the CTV of the internal mammary nodal region, ipsilateral brachial plexus, left anterior descending coronary artery, and right coronary artery were 0.38, 0.21, 0.29, and 0.18, respectively, suggesting marked variations. In addition, marked under- and overoutlining variations were identified on 4 slices of CTVsc + axIII + axII on slice-by-slice analysis. CONCLUSIONS: There were residual contouring variations despite a detailed protocol being provided, confirming the importance of pretrial QA in RT and highlighting the need for education and consideration of a real-time central review of the target delineation before the trial participants begin RT.

Benefits of combination therapy with immune checkpoint inhibitors and predictive role of tumour mutation burden in hepatocellular carcinoma: A systematic review and meta-analysis.

OBJECTIVES: To investigate the clinical benefits of combination therapy with immune checkpoint inhibitors (ICIs) and best combination regimen for people with advanced hepatocellular carcinoma (HCC) and to explore the predictive performance of tumour mutation burden (TMB). METHODS: We conducted a systematic literature search to identify clinical trials. Meta-analysis and subgroup analyses were performed to estimate the benefits of combination regimens with PD-1/PD-L1 inhibitors for patients with advanced HCC and compare the effectiveness of PD-1/PD-L1 inhibitors and sorafenib as first-line therapy. Individualized analysis and Kaplan-Meier were used to assess the prognostic value of TMB. RESULTS: A total of 29 studies with 5396 patients were included. ICIs' combination therapy had higher ORR (26 % vs 15 %) and DCR (73 % vs 55 %), longer PFS (5.5 vs 3.1 months) and OS (15.9 vs 12.6 months) compared to monotherapy. Anti-PD-1/PD-L1 agents provided improved ORR, DCR, PFS and OS compared to sorafenib. The overall ORs of ORR and DCR in subgroup analysis were 3.49 (95 % CI 2.36-5.17, p < 0.01) and 1.60 (95 % CI 1.15-2.21, p < 0.01). The overall HRs of PFS and OS were 0.68 (95 % CI 0.48-0.96, p = 0.03) and 0.73 (95 % CI 0.62-0.85, p < 0.01). PD-1/PD-L1 inhibitors plus anti-VEGF agents had an advantage in DCR (0.80 vs 0.48, meta-regression =  - 0.32, P < 0.001), but an equal ORR (0.29 vs 0.26) compared to dual immune checkpoint inhibitors. The total OS in Dua-ICIs were 16.5 months (95 % CI 14.2-18.7), yet not reached in the major studies of ICI plus anti-VEGF regimen. In individualized analysis, the 1-year OS was superior for patients who had high-TMB (>10, mutations/Mb) than moderate-TMB (1-10, mutations/Mb; 28 % vs 15 %, P = 0.025). CONCLUSION: Immune checkpoint inhibitors' combination therapy improved clinical outcomes in the management of advanced hepatocellular carcinoma. However, the overall objective response rate still did not exceed 30%. PD-1/PD-L1 inhibitors plus anti-angiogenic agents and dual immunotherapy provided significantly increased survival over sorafenib, which also pose new challenges for future research, and more appropriate and guided control regimens are required. Also, TMB may be a promising prognostic biomarker for immunotherapy in HCC. However, the validation of prospective and large sample studies is needed.

Separation of Uranium and Thorium for 230Th-U Dating of Submarine Hydrothermal Sulfides.

The age of a submarine hydrothermal sulfide is a significant index for estimating the size of hydrothermal ore deposits. Uranium and thorium isotopes in the samples can be separated for 230Th-U dating. This article presents a method to purify and separate U and Th isotopes in submarine hydrothermal sulfide samples. Following this technique, the separated U and Th fractions can meet measuring requirements by multi-collector inductively coupled plasma mass spectrometry (MC-ICPMS). The age of the hydrothermal sulfide sample can be calculated by measuring the present-day activity ratios of 230Th/238U and 234U/238U. A super clean room is necessary for this experiment. Cleaned regents and supplies are used to reduce the contamination during the sample processes. Balance, hotplate, and centrifuge are also used. The sulfide sample is powdered for analysis and less than 0.2 g sample is used. Briefly, the sample is weighed, dissolved, added to 229Th-233U-236U double spike solution, Fe co-precipitated, and separated on an anion-exchange resin extraction column. Approximately 50 ng U is consumed for 230Th-U dating of sulfides sample by MC-ICPMS.

Increased expression of O-GlcNAc transferase (OGT) is a biomarker for poor prognosis and allows tumorigenesis and invasion in colon cancer.

Recent studies suggest that Elevated O-GlcNAcylation by increased O-GlcNAc transferase (OGT) and/or decreasing O-GlcNAcase (OGA) levels is associated with cancer initiation, progression, invasion, and metastasis. However, the function of OGT in colon cancer tumorigeneses remains unclear. Here, we showed OGT expression is elevated in human colon cancer tissue compared with adjacent normal tissue, and cases with higher OGT expression had shorter survival time. Additionally, OGT mRNA expression was positively correlated with pathologic TNM stage from TCGA public database. Finally, we found knock-down of OGT expression by RNA interference inhibits cell proliferation, migration and invasion in colon cancer cell lines. Taken together, this study imply that elevated OGT expression had an important function in colon cancer formation and progression, and OGT may be a valuable prognostic factor and therapeutic target.

Phase I study of icotinib, an EGFR tyrosine kinase inhibitor combined with IMRT in nasopharyngeal carcinoma.

BACKGROUND: Epidermal growth factor receptor (EGFR) is a new target for nasopharyngeal carcinoma (NPC) therapy. This prospective phase I study sought to determine the safety and recommended phase II dose of icotinib, a novel highly selective oral EGFR tyrosine kinase inhibitor, in combination with intensity-modulated radiotherapy (IMRT) in patients with NPC. METHODS: Eligible patients with NPC received escalating doses of icotinib during IMRT. We treated six patients at a particular dose level until the maximum tolerated dose (MTD) was determined. The starting dose was 125 mg, once-daily and the dose was escalated to another level 125 mg, twice- and thrice- daily, until dose-limiting toxicity (DLT) occurred in two or more patients at a dose level. Expression and mutation analysis of EGFR were performed in all cases. RESULTS: A total of twelve patients were enrolled. Three patients experienced DLT (250 mg/day cohort) and MTD was 125 mg/day. Mucositis toxicity appears to be the major DLT. While EGFR expression in tumor tissue was detected in 75% (9/12) patients, EGFR mutation was detected in 16.67% (1/6) patients in 125 mg/day cohort, and 50% (3/6) in 250 mg/day cohort. CONCLUSION: The combination of icotinib (125 mg/day) and IMRT in patients with locally NPC had an acceptable safety profile and was well tolerated.

Nimotuzumab abrogates acquired radioresistance of KYSE-150R esophageal cancer cells by inhibiting EGFR signaling and cellular DNA repair.

BACKGROUND: Acquired radioresistance of cancer is common after repeated irradiation and often leads to treatment failure. This study aimed to examine the effects of nimotuzumab on acquired radioresistance in human esophageal carcinoma cells and to investigate its underlying mechanisms. METHODS: The radioresistant human esophageal carcinoma cell line KYSE-150R was generated by using fractionated irradiation. KYSE-150R cells were pretreated with or without nimotuzumab before ionizing radiation. Cell growth and colony formation were measured to quantitate the effects of radiation. The γ-H2AX foci assay was employed to determine cellular DNA-repairing capacity. The phosphorylation of key molecules involved in the epidermal growth factor receptor (EGFR) signaling pathway and cellular DNA repair was measured by Western blot analysis. RESULTS: Nimotuzumab enhanced radiation-induced inhibition on cell growth and clonogenic survival in KYSE-150R cells. The average number of γ-H2AX foci increased in the irradiated cells treated with nimotuzumab. Nimotuzumab inhibited phosphorylation of the EGFR and its downstream molecules AKT and ERK. Phosphorylation of the DNA repair-related proteins DNA-PKcs, ATM, and RAD51 was also inhibited by nimotuzumab. CONCLUSIONS: These results indicate that nimotuzumab can inhibit key cancer survival mechanisms, the EGFR signaling pathway, and DNA repair and thereby reverse acquired radioresistance in KYSE-150R cell line.

Fasting blood glucose, but not 2-h postload blood glucose or HbA1c, is associated with mild decline in estimated glomerular filtration rate in healthy Chinese.

BACKGROUND: The association of blood glucose [including fasting blood glucose (FBG), 2-h postload blood glucose, and glycated hemoglobin (HbA1c)] with the risk of a mild decline in the estimated glomerular filtration rate (eGFR) in healthy subjects was unclear. The aim of the study was to investigate this association in middle-aged and elderly healthy Chinese. METHODS: The study included 1,112 healthy Chinese who were ≥40 years old, and all the subjects were divided into two groups based on FBG value of 5.6 mmol/L. A mildly reduced eGFR was defined as 60-90 mL/min/1.73 m2. Multiple linear or logistic regression analysis was used to estimate the association of blood glucose with eGFR and the risk of a mildly reduced eGFR, respectively. A generalized additive model was used to explore a possible nonlinear relationship between FBG and eGFR. RESULTS: FBG was significantly associated with decreased eGFR and increased risk of a mildly reduced eGFR independent of age, gender, body mass index, waist circumference, systolic blood pressure (BP), diastolic BP, triglyceride, high-density lipoprotein cholesterol, fasting insulin, smoking, and drinking. Additionally, FBG and eGFR showed a nonlinear association (P<0.001). Interestingly, the group with FBG≥5.6 mmol/L significantly increased the risk of a mildly reduced eGFR (OR 2.06, P=0.001) after multivariable adjustment. CONCLUSIONS: FBG is closely associated with decreased eGFR and increased risk of a mildly reduced eGFR. The ADA criteria using FBG=5.6 mmol/L instead of 6.1 mmol/L as cutoff point to define prediabetes are more suitable for the prevention of diabetes and related kidney disease.

Fluctuation between fasting and 2-H postload glucose state is associated with glomerular hyperfiltration in newly diagnosed diabetes patients with HbA1c < 7%.

OBJECTIVE: To investigate whether fluctuations between the fasting and 2-h postload glucose ([2-hPBG]-fasting blood glucose [FBG]) states are associated with glomerular hyperfiltration (GHF) in middle-aged and elderly Chinese patients with newly diagnosed diabetes. DESIGN AND METHODS: In this study, we included 679 newly diagnosed diabetes patients who were ≥ 40 years old. All the subjects were divided into two groups; those with HbA1c<7% and ≥ 7%. The Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation was used to estimate the glomerular filtration rate (GFR). GHF was defined as an eGFR ≥ the 90th percentile. First, a multiple linear regression analysis was used to estimate the association of 2-hPBG-FBG with eGFR. Then, a generalized additive model was used to explore the possible nonlinear relationship between 2-hPBG-FBG and eGFR. Next, the 2-hPBG-FBG values were divided into four groups as follows: 0-36, 36-72, 72-108 and ≥ 108 mg/dl. Finally, a multiple logistic regression analysis was used to investigate the association of 2-hPBG-FBG with the risk of GHF. RESULTS: For the group with HbA1c<7%, the eGFR and the percentage of GHF were significantly higher compared with the group with HbA1c ≥ 7%. After adjusting for age, gender, body mass index (BMI), systolic blood pressure (BP), diastolic BP, fasting insulin, cholesterol, triglycerides, smoking, drinking and glycated hemoglobin (HbA1c), 2-hPBG-FBG was significantly associated with increased eGFR and an increased risk of GHF (the GHF risk increased by 64.9% for every 36.0 mg/dl [2.0 mmol/L] 2-hPBG-FBG increase) only in those patients with HbA1c<7%. Additionally, 2-hPBG-FBG and eGFR showed a nonlinear association (P<0.001). CONCLUSIONS: Increased fluctuations between the fasting and 2-h postload glucose states are closely associated with increased eGFR and an increased risk of GHF in newly diagnosed diabetes patients with HbA1c<7%.

Triglyceride levels are closely associated with mild declines in estimated glomerular filtration rates in middle-aged and elderly Chinese with normal serum lipid levels.

OBJECTIVE: To investigate the relationship between lipid profiles [including total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C)] and a mild decline in the estimated glomerular filtration rate (eGFR) in subjects with normal serum lipid levels. DESIGN AND METHODS: In this study, we included 2647 participants who were ≥ 40 years old and had normal serum lipid levels. The Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation was used to estimate the GFR. A mildly reduced eGFR was defined as 60-90 mL/min/1.73 m(2). First, multiple linear regression analysis was used to estimate the association of lipid profiles with the eGFR. Then, the levels of each lipid component were divided into four groups, using the 25th, 50th and 75th percentiles as cut-off points. Finally, multiple logistic regression analysis was used to investigate the association of different lipid components with the risk of mildly reduced eGFR. RESULTS: In the group with a mildly reduced eGFR, TG and LDL-C levels were significantly increased, but HDL-C levels were significantly decreased. After adjusting for age, gender, body mass index (BMI), systolic blood pressure (SBP), glycated hemoglobin (HbA1c), smoking and drinking, only TC and TG were independently related to the eGFR. Additionally, only TG showed a linear relationship with an increased risk of a mildly reduced eGFR, with the highest quartile group (TG: 108-150 mg/dl [1.22-1.70 mmol/L]) having a significantly increased risk after adjusting for the above factors. CONCLUSIONS: Triglyceride levels are closely associated with a mildly reduced eGFR in subjects with normal serum lipid levels. Dyslipidemia with lower TG levels could be used as new diagnostic criteria for subjects with mildly reduced renal function.

Metabolic abnormalities, but not obesity, contribute to the mildly reduced eGFR in middle-aged and elderly Chinese.

BACKGROUND: The role of obesity as a determinant of kidney dysfunction has not reached an agreement and the underlying reason may be due to the heterogeneity of obese phenotypes. The aim of the study was to explore the associations of different obese phenotypes with the change of estimated glomerular filtration rate (eGFR) and the roles of obesity and metabolic abnormalities in this association. METHODS: eGFR was calculated in 8,586 participants (≥ 40 years old). eGFR 60-90 mL/min/1.73 m(2) was defined as the mildly reduced eGFR. Multiple logistic regression analysis was used to determine odds ratios (ORs) for mildly reduced eGFR in the metabolically healthy obese (MHO), metabolically abnormal non-obese (MANO) and metabolically abnormal obese (MAO) groups, using the metabolically healthy non-obese (MHNO) subjects as the reference group. Meanwhile, the associations of body mass index (BMI), waist circumference (WC) and metabolic abnormalities (including hypertension, hyperglycemia and dyslipidemia) with the risk of mildly reduced eGFR were also investigated. RESULTS: The proportion of MHNO, MHO, MANO and MAO subjects was 8.3, 17.1, 10.1 and 64.5 %, respectively. Increased ORs were observed in MANO (OR 1.51, P = 0.014) and MAO (OR 1.47, P = 0.015) groups, after adjusting for age, gender, smoking, drinking, BMI and WC. When further adjusting for metabolic abnormalities, MANO (OR 1.24, P = 0.247) and MAO (OR 1.17, P = 0.366) subjects would not present increased risk of mildly reduced eGFR any more. Oppositely, fasting insulin (OR 1.03, P < 0.001), hyperglycemia (OR 1.25, P = 0.002) and dyslipidemia (OR 1.25, P = 0.002), but not hypertension, BMI and WC, significantly increased the risk of mildly reduced eGFR. CONCLUSIONS: Metabolic abnormalities, but not simple obesity, may contribute to the mildly reduced eGFR in middle-aged and elderly Chinese.

Fluctuation between fasting and 2-H postload glucose state is associated with chronic kidney disease in previously diagnosed type 2 diabetes patients with HbA1c ≥ 7%.

OBJECTIVE: To investigate how the glucose variability between fasting and a 2-h postload glucose state (2-h postload plasma glucose [2hPG]-fasting plasma glucose [FPG]) is associated with chronic kidney disease (CKD) in middle-aged and elderly Chinese patients previously diagnosed with type 2 diabetes. DESIGN AND METHODS: This cross-sectional study included 1054 previously diagnosed type 2 diabetes patients who were 40 years of age and older. First, the subjects were divided into two groups based on a glycated hemoglobin (HbA1c) value of 7%. Each group was divided into two subgroups, with or without CKD. The Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation was used to estimate the glomerular filtration rate (GFR). CKD was defined as eGFR<60 mL/min/1.73 m2. Multiple linear regression analysis was used to estimate the association between the 2hPG-FPG and eGFR. The 2hPG-FPG value was divided into four groups increasing in increments of 36 mg/dl (2.0 mmol/L): 0-72, 72-108, 108-144 and ≥144 mg/dl, based on the quartiles of patients with HbA1c levels ≥7%; then, binary logistic regression analysis was used to investigate the association between 2hPG-FPG and the risk of CKD. RESULTS: In the patients with HbA1c levels ≥7%, the 2hPG-FPG was significantly associated with decreased eGFR and an increased risk of CKD independent of age, gender, body mass index (BMI), systolic blood pressure (BP), diastolic BP, smoking, and drinking, as well as fasting insulin, cholesterol, triglyceride, and HbA1c levels. The patients with 2hPG-FPG values ≥144 mg/dl showed an increased odds ratio (OR) of 2.640 (P = 0.033). Additionally, HbA1c was associated with an increased risk of CKD in patients with HbA1c values ≥7%. CONCLUSIONS: The short-term glucose variability expressed by 2hPG-FPG is closely associated with decreased eGFR and an increased risk of CKD in patients with poor glycemic control (HbA1c≥7%).

Bmi-1 confers adaptive radioresistance to KYSE-150R esophageal carcinoma cells.

Radiotherapy (RT) is a major modality of cancer treatment. However, tumors often acquire radioresistance, which causes RT to fail. The exact mechanisms by which tumor cells subjected to fractionated irradiation (FIR) develop an adaptive radioresistance are largely unknown. Using the radioresistant KYSE-150R esophageal squamous cell carcinoma (ESCC) model, which was derived from KYSE-150 parental cells using FIR, the role of Bmi-1 in mediating the radioadaptive response of ESCC cells to RT was investigated. The results showed that the level of Bmi-1 expression was significantly higher in KYSE-150R cells than in the KYSE-150 parental cells. Bmi-1 depletion sensitized the KYSE-150R cells to RT mainly through the induction of apoptosis, partly through the induction of senescence. A clonogenic cell survival assay showed that Bmi-1 depletion significantly decreased the radiation survival fraction in KYSE-150R cells. Furthermore, Bmi-1 depletion increased the generation of reactive oxygen species (ROS) and the expression of oxidase genes (Lpo, Noxo1 and Alox15) in KYSE-150R cells exposed to irradiation. DNA repair capacities assessed by γ-H2AX foci formation were also impaired in the Bmi-1 down-regulated KYSE-150R cells. These results suggest that Bmi-1 plays an important role in tumor radioadaptive resistance under FIR and may be a potent molecular target for enhancing the efficacy of fractionated RT.

Changes of the transverse diameter and volume and dosimetry before the 25th fraction during the course of intensity-modulated radiation therapy (IMRT) for patients with nasopharyngeal carcinoma.

To quantify changes of the transverse diameter and volume and dosimetry, and to illustrate the inferiority of non-replanning during intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC) patients. Fifty-three NPC patients who received IMRT in 33 fractions were enrolled in this prospective trial. Before the 25th fraction, a new simulation computed tomography (CT) scan was acquired for all patients. The dose-volume histograms of the phantom plan were compared with the initial plan. Significant reduction of the transverse diameter of the nasopharyngeal, the neck, and 2 parotid glands volume was observed on second CT compared with the first CT (mean reduction 7.48 ± 4.45 mm, 6.80 ± 15.14 mm, 5.70 ± 6.26 mL, and 5.04 ± 5.85 mL, respectively; p < 0.01). The maximum dose and V-40 of the spinal cord, mean dose, and V30 of the left and right parotid, and V-50 of the brain stem were increased significantly in the phantom plan compared with the initial plan (mean increase 4.75 ± 5.55 Gy, 7.18 ± 10.07%, 4.51 ± 8.55 Gy, 6.59 ± 17.82%, 5.33 ± 8.55 Gy, 11.68 ± 17.11% and 1.48 ± 3.67%, respectively; p < 0.01). On the basis of dose constraint criterion in the RTOG0225 protocol, the dose of the normal critical structures for 52.83% (28/53) of the phantom plans were out of limit compared with 1.89% (1/53) of the initial plans (p < 0.0001). Because of the significant change in anatomy and dose before the 25th fraction during IMRT, replanning should be necessary during IMRT with NPC.