Determination of EGFR-overexpressing tumor cells by magnetic gold-decorated graphene oxide nanocomposites based impedance sensor.

This work developed an electrochemical impedance spectroscopy (EIS) sensor for detection of EGFR (epidermal growth factor receptor)-overexpressing tumor cell and preliminary estimation of EGFR expression. Here, EGFR antibodies as the specific antibodies for cancer cells were conjugated on magnetic gold-decorated graphene oxide nanocomposites, which were used to capture the EGFR-overexpressing tumor cells. The magnetically responsive tumor cells were enriched and immobilized on a magnetic glassy carbon electrode (mGCE) surface, leading to increased electron-transfer resistance (Ret) utilized for determination of cells and preliminary evaluation of EGFR expression level. This strategy enables the enrichment, fixation and detection of tumor cells to be accomplished in a facile way. An excellent linearity in the range of 2.0 × 102 - 3.0 × 105 cell mL-1 with the detection limit of 152 cell mL-1 for MDA-MB-231 cells was obtained. Investigation on the expression levels of EGFR on various types of cells was conducted. MDA-MB-231 cells showed a distinctly higher EGFR expression, compared with MHCC97-L and L02 cells, providing the possibility for the EGFR-targeted therapy of the tumors. It is expected that the proposed sensor has the potential to be applied for cancer monitoring.

Joint Replacement Surgeries Can Be Safely Performed in HIV Patients.

Whether joint replacement surgery can be performed safely on HIV patients is still a matter of debate. This study aimed to report the surgical efficacy and complications of joint replacement surgery in HIV patients. A total of 21 HIV patients and 27 non-HIV patients who underwent arthroplasties in our hospital were retrospectively reviewed. The 21 HIV patients received 29 joint replacement surgeries including 27 cases of total hip arthroplasty (THA) and 2 cases of total knee arthroplasty (TKA). The non-HIV patients received 16 THA, 10 TKA, and 3 unicompartmental arthroplasty (UKA). The length of hospital stay of HIV patients was significantly lower than that of non-HIV patients. At the last follow-up, there were no significant complications both in the HIV group and the non-HIV groups. No medical staff had any occupational exposure. We concluded that joint replacement surgery in HIV patients is safe and effective. Optimization of patients is key to treatment success. Strictly following the standardized protection protocol can prevent the risk of occupational exposure.

Quercetin-Loaded Ceria Nanocomposite Potentiate Dual-Directional Immunoregulation via Macrophage Polarization against Periodontal Inflammation.

Macrophage polarization toward M1 phenotype (pro-inflammation) is closely associated with the destructive phase of periodontal inflammation. Nanoceria is verified to inhibit M1 polarization of macrophages by the favorable ability of reactive oxygen species (ROS) scavenging. However, the function of nanoceria on macrophage polarization toward M2 phenotype (anti-inflammation) in reparative phase of periodontal inflammation is quite limited. In this work, by introducing an antioxidant drug quercetin onto nano-octahedral ceria, synergistic and intense regulation of host immunity against periodontal disease is realized. Such nanocomposite can control the phenotypic switch of macrophages by not only inhibition of M1 polarization for suppressing the damage in the destructive phase but also promotion of M2 polarization for regenerating the surrounding tissues in reparative phase of periodontal disease. As-prepared nanocomposite can effectively increase the M2/M1 ratio of macrophage polarization in inflammatory cellular models by lipopolysaccharide stimulation. More importantly, the nanocomposite also exerts an improved therapeutic potential against local inflammation by significant downregulation of pro-inflammatory cytokines and upregulation of anti-inflammatory cytokines in an animal model with periodontal inflammation. Therefore, this newly developed nanomedicine is efficient in ROS scavenging and driving pro-inflammatory macrophages to the anti-inflammatory phenotype to eliminate inflammation, thereby providing a promising candidate for treating periodontal inflammation.

Drug delivery based pharmacological enhancement and current insights of quercetin with therapeutic potential against oral diseases.

The heavy burden of oral diseases such as oral cancers, dental caries, periodontitis, etc. and their consequence on the patient's quality of life demonstrated an urgent demand for developing effective therapeutics. Quercetin as a natural derived flavonoid, could be utilized in the therapeutic formulation of various diseases such as diabetes, breast cancer and asthma, owing to its prominent pharmacological values. In the last decade, the applications of quercetin as a natural compound in oral treatment have attracted increasing interest due to its multifunction including antioxidant, antibacterial, anti-inflammatory and antineoplastic activities. Besides, considering the low bioavailability of quercetin, great efforts have been made in its drug delivery systems to address the problem of limited application. Therefore, this review summarized the cutting-edge researches on versatile effects and enhanced bioavailability of quercetin resulting from innovative drug delivery systems, particularly focused on its potential against oral diseases. The application of quercetin would provide novel and promising therapeutic approach for clinical treatment, promoting the development of global dental public health.

FOXO1 Mediates Advanced Glycation End Products Induced Mouse Osteocyte-Like MLO-Y4 Cell Apoptosis and Dysfunctions.

Osteocyte plays an essential role in bone metabolism by regulating osteoblast and osteoclast activities. Dysfunction or apoptosis of osteocyte will severely endanger the bone homeostasis and result in bone diseases such as osteoporosis. Osteoporosis has been considered as one of the diabetes complications; however, the mechanism is still to be discovered. Advanced glycation end products (AGEs), as the main pathogenic factor of diabetes mellitus, have the capacity to induce osteocyte apoptosis thus sabotaging bone homeostasis. Here, we examined the role of AGE during osteocyte apoptosis and how this effect would affect osteocyte's regulation of osteoblast and osteoclast. Mouse osteocyte-like MLO-Y4 cells were used to study the properties of osteocyte and to examine its biological and pathological function. MTT assay and Annexin V assay showed that AGE significantly induce MLO-Y4 cell apoptosis. qPCR and Western blot results have shown that AGE upregulates proapoptotic gene p53 and its downstream target gene Bax, which leads to enhanced activation of caspase-3, thus inducing apoptosis in MLO-Y4 cells. Increased expression of sclerostin and RANKL in osteocytes has shown that AGE induces osteocyte dysfunction thus severely damaging the bone homeostasis by decreasing osteoblast and increasing osteoclast activities. Furthermore, the role of the transcription factor FOXO1, which is intensely associated with apoptosis, has been determined. Western blot has shown that AGE significantly decreases Akt activities. Immunofluorescence has shown that AGE promotes FOXO1 nuclei localization and enhances FOXO1 expression. Silencing of FOXO1 suppressed AGE-enhanced apoptosis; mRNA and protein expressions of cleaved caspase-3, sclerostin, and RANKL were downregulated as well. Moreover, exogenous FOXO1 increased caspase-3 mRNA levels and caspase-3 transcriptional activity. Lastly, ChIP assay has established the capacity of FOXO1 binding directly on the caspase-3, sclerostin, and RANKL promoter region in AGE environment, providing the mechanism of the AGE-induced osteocyte apoptosis and dysfunction. Our results have shown that FOXO1 plays a crucial role in AGE-induced osteocyte dysfunction and apoptosis through its regulation of caspase-3, sclerostin, and RANKL. This study provides new insight into diabetes-enhanced risk of osteoporosis given the critical role of AGE in the pathogenesis of diabetes and the essential part of osteocyte in bone metabolism.

One-Stage Positron Emission Tomography and Magnetic Resonance Imaging to Assess Mesenchymal Stem Cell Survival in a Canine Model of Intervertebral Disc Degeneration.

Intervertebral disc degeneration (IVDD) is a major health problem. Although mesenchymal stem cells (MSCs) have been used to promote IVD regeneration, the actual survival time of implanted MSCs in IVDs has never been studied noninvasively and continuously in vivo. To investigate survival of implanted MSCs in vivo, this study used a canine model of degenerated IVD and MSCs transfected with a mutant herpes simplex type-1 virus thymidine kinase and labeled with magnetic iron oxide nanoparticles (MION). One-stage positron emission tomography (PET) and magnetic resonance (MR) imaging were carried out 3 days and 2 weeks, 3 weeks, and 4 weeks after implantation of MSCs into IVDs with surgically induced degeneration. Pfirrmann disc degeneration grade determined from the MR images indicated that the repair progress of degenerated IVD stopped 3 weeks after MSC implantation. Meanwhile, MION signal strength, signal contrast ratio (%), and low signal area (mm2) did not change significantly from that seen 3 days after cell implantation until 4 weeks [751.43 (4 weeks) ±52.67 (3 days) vs. 225.34 ± 35.62; 47.37 ± 5.01 vs. 85.37 ± 10.54; 1.78 ± 0.31 vs. 5.29 ± 1.35; P < 0.01, respectively]. Accumulation of the PET reporter probe, 9-(4-[18F]-fluoro-3-hydroxymethylbutyl)-guanine, was dramatically decreased at 3 weeks after MSC implantation. These results demonstrated that MSCs could survive no more than 3 weeks after implantation into IVDs with surgically induced degeneration, suggesting that MSCs could contribute to IVD repair for the first 3 weeks after implantation. The results also indicate that PET imaging could be used reliably to quantify the survival of implanted MSCs, whereas MION with MR imaging would likely be unsuitable for long-term tracking of MSCs in IVDs.

Urinary metabolomics reveals the therapeutic effect of HuangQi Injections in cisplatin-induced nephrotoxic rats.

The side effects of cisplatin (CDDP), notably nephrotoxicity, greatly limited its use in clinical chemotherapy. HuangQi Injections (HI), a commonly used preparation of the well-known Chinese herbal medicine Astragali radix, appeared to be promising treatment for nephrotoxicity without compromising the anti-tumor activity of CDDP. In this study, the urinary metabolomics approach using liquid chromatography time of flight mass spectrometry (LC-TOF/MS) was developed to assess the toxicity-attenuation effects and corresponding mechanisms of HI on CDDP-exposed rats. As a result, successive administration of HI significantly recovered the decline of body weight and downregulated the abnormal increase of serum creatinine and urea. HI partly restored the CDDP-induced alteration of metabolic profiling back into normal condition. Totally 43 toxicity-attenuation potential biomarkers were screened and tentatively identified, which were involved in important metabolic pathways such as amino acid metabolism, TCA cycle, fatty acid metabolism, vitamin B6 metabolism and purine metabolism. The results clearly revealed that HI could alleviate CDDP-induced nephrotoxicity and improve the disturbed metabolic balance induced by repeated CDDP exposure. The present study provided reliable evidence for the protective effect of HI on CDDP-induced toxicity with the multi-target pharmacological characteristics.

Determination of Cadmium in Shrimp and Shell Fish Samples by Coprecipitation Enrichment with Mn(â ¡)-5-Br-PADAP Flame Atomic Absorption Spectrometry.

A separation/preconcentration procedure with coprecipitation has been proposed for the flame atomic absorption spectrometric (FAAS) determination of cadmium at trace level in food and environmental samples. Manganese(Ⅱ) was used as a carrier which chelated with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol to detect the content of trace cadmium in shrimp and shell fish samples with flame atomic absorption spectrometry for the first time. The precipitate could be easily dissolved with concentrated nitric acid. The optimum coprecipitation of this new method including the amount of reagent, amount of manganese(Ⅱ), the pH, and the standing time of the precipitate had been confirmed for the quantitative recoveries of the analytes. The effect of matrix ions and the interference of co-existing ions were also evaluated. Under the experimental conditions established by the optimization step, the system of Mn(Ⅱ)-5-Br-PADAP was able to overcome the matrix interference which showed the effect of separation and enrichment well. The linear range of cadmium content was determined to be 0.1～1.0 mg·L-1. The sensitivity and the relative standard deviation(RSD) were found 0.147(mg·L-1)-1, 0.73%, respectively. The optimum procedure allows the determination of cadmium with limit of detection of 4.27 µg·L-1. The complexity of preprocessing was determined by the complexity of food samples. So the differences of cadmium content in the samples between the direct determination with atomic absorption spectrometry and the measurement after coprecipitation were examined, which providedevidences for the superiority of the system again. Cadmium in shell fish and shrimp samples were 1.85 mg·kg-1 and 1.74 mg·kg-1, which in line with international standards of the Codex Alimentarius Commission(CAC). The credibility of the method was evaluated by standard additional method and recovery experiments. The standard addition recoveries of sample and RSDs of the method were in the range of 99.9%～100.3% and 0.15%～0.83%. The results of recovery experiment showed that the presented coprecipitation procedure had good repetition, high accuracy. In addition, with the method, we could draw conclusions that the experiments were simple and rapid. The developed method described in the literature was successfully applied for the determination of trace cadmium in shrimp and shell fish samples with satisfactory results.

[Effect of assisted anesthesia of auricular point magnetic sticking on postoperative recovery of gynecological surgery].

OBJECTIVE: To explore an optimum method on postoperative recovery of general-anesthesia gynecological laparoscopic surgery. METHODS: One hundred and twenty cases of gynecological laparoscopic surgery were randomly divided into three groups, 40 cases in each one. The sensitive points of bilateral Shenmen (TF4), Zigong (TF2), Wei (CO4), Dachang (CO7) and so on were selected in all three groups one night before the surgery. The sticking with magnetic beads was applied in the group A. The magnetic beads were in-pair stuck at front-back corresponding location of both ears in the group B. The same-appearance plaster was put at the corresponding acupoints in the group C. The intubation anesthesia was applied in all three groups and postoperative recovery of gynecological laparoscopic surgery was observed. RESULTS: The postoperative visual analogue scale (VAS) was (1.77 +/- 1.65) in the group A and (1.80 +/- 1.96) in the group B, which was both lower than (2.62 +/- 1.46) in the group C (both P < 0.01). The occurrence rate of nausea and vomiting was 25.0% (9/36) in the group A and 20.0% (8/40) in the group B, which was both lower than 50.0% (19/38) in the group C (both P < 0.01). The recovery of borborygmus on postoperative 1st and 3rd day in the group A and B was faster than that in the group C (both P < 0.05), while time of fart and defecation in the group A and B was earlier than that in the group C (both P < 0.05). The score of state-trait anxiety inventory in the group A and B was lower than that in the group C (both P < 0.05), but the differences of each item between the group A and B were not obvious (both P > 0.05). CONCLUSION: The auricular point sticking could support analgesia of general-anesthesia gynecological laparoscopic surgery, which could relieve anxiety mood, reduce occurrence of nausea and vomiting and improve function of stomach and intestine to benefit postoperative recovery. However, the effect of in-pair sticking of auricular point with magnetic beads at front-back acupoints was not obviously strengthened.

[Observation on the anesthesia effect of general anesthesia assisted by auricular-plaster therapy in gynecological surgery].

OBJECTIVE: To observe the impacts on anesthetic dosage and postoperative pain under general anesthesia assisted by auricular-plaster therapy. METHODS: One hundred and twenty cases of gynecological laparoscopic surgery were randomized into three groups, 40 cases in each one. In auricular point group A, the magnetic beads were stick on the auricular points which were Shenmen (TF4), Lung (fei, CO14), Spleen (pi,CO13), Stomach (wei, CO4), Large Intestine (dachang, CO7), Adrenal (shenshangxian, TG2 (P)), Endocrine (neifenmi, CO18), Uterus and Pelvic Cavity(penqiang, TF5) etc. In auricular point group B, the magnetic beads were stick on the preauricular points and the corresponding retroauricular points of the ears. In a placebo group, the plasters of the same appearance were stick on the corresponding auricular points of the ears. The anesthetic method was same in three groups and the anesthesia effect were assessed and compared among the three groups. RESULTS: In surgery, the dosage of sufentanil, the narcotic analgesic was (22.08 +/- 7.11) microg in auricular point group A and was (20.19 +/- 7.21) microg in auricular point group B, which were reduced as compared with (26.13 +/- 9.04) microg in the placebo group (both P < 0.05). The difference in the dosage of propofol, the anesthetic was not significant statistically among three groups (P > 0.05). On the 1st day after surgery, the score of pain visual analogue scale (VAS) was (1.77 +/- 1.65) in auricular point group A and was (1.80 +/- 1.96) in auricular point group B, which were reduced as compared with (2.62 +/- 1.46) in the placebo group (both P < 0.05). Before surgery, the serum beta-endorphin (beta-EP) was (198.8 +/- 124.1) pg/mL in auricular point group A and was (207.2 +/-102.7) pg/mL in auricular point group B, which were higher apparently as compared with (143.6 +/- 87.1) pg/mL in the placebo group (both P < 0.05). The differences in the above indices were not significant statistically between the two auricular point groups. CONCLUSION: The auricular-plaster therapy reduces the dosage of anesthetic, alleviates postoperative pain and acts on tranquilization and analgesia. The effect is not intensified apparently in the treatment for the magnetic beads sticking on both preauricular points and the corresponding retroauricular points of the ears.

[Creation of tissue engineered cartilage with internal support].

OBJECTIVE: To test the hypothesis that tissue-engineered cartilage can be bioincorporated with a nonreactive, permanent endoskeletal scaffold. METHODS: Chondrocytes obtained from swine articular were seeded onto polyglycolic acids(PGA) scaffold which was incorporated with high-density polyethylene (Medpor). After cultured in vitro for two weeks,the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. Six weeks later,the newly formed cartilage prosthesis was harvested, and a small part of sample was evaluated by gross view, histology, type II collagen immunohistochemistry and biochemistry. PGA scaffold seeded with cells as the control group. RESULTS: The newly formed cartilage was very similar to normal cartilage in both gross view and histology, and jointed Medpor tightly. The center of control group was hollow. CONCLUSION: This pilot technique combining tissue engineering with a permanent success in creating cartilage without "hollow" phenomenon. biocompatible endoskeleton demonstrated

[Effect of previous analgesia of scalp acupuncture on post-operative epidural morphine analgesia in the patient of intestinal cancer].

OBJECTIVE: To probe the effect of previous analgesia of scalp acupuncture. METHODS: Sixty cases of radical operation of intestinal cancer were randomly divided into 2 groups: a scalp acupuncture plus epidural analgesia (scalp acupuncture group) and an epidural analgesia group, 30 cases in each group. The scalp acupuncture group received scalp acupuncture from 20 min before operation to the end of the operation, and epidural analgesia at the end of the operation. The epidural analgesia group only received epidural analgesia at the end of the operation. Recovery of VAS scores, BCS scores and gastrointestinal function after operation were investigated to probe into effects of scalp acupuncture on epidural morphine analgesia after operation in the patient of intestinal cancer. RESULTS: The VAS scores at 6 h, 12 h, 24 h and 48 h in the scalp acupuncture group were lower than the epidural analgesia group, with significant differences at 6 h and 12 h between the two groups (P < 0.05). BCS scores at the 4 time points in the scalp acupuncture group were lower than the epidural analgesia group with significant differences between the two groups (all P < 0.05). The scalp acupuncture group in recovery of bowel sound, exsufflation from the anus in the scalp acupuncture group was faster than the epidural analgesia group (P < 0.01). CONCLUSION: Scalp acupuncture has a certain previous analgesic effect, reducing discomfort of the patient after operation and promoting the recovery of gastrointestinal functions.

[Effects of scalp acupuncture on regulative ability of oxidative stress reaction at operation stage].

OBJECTIVE: To study on regulative action of scalp acupuncture on oxidative stress reaction at operation stage. METHODS: Fifty two cases selected for radical operation of intestinal cancer were randomly divided into a scalp acupuncture plus general anesthesia (scalp acupuncture group) and a simple general anesthesia group (general anesthesia group), 26 cases in each group. The scalp acupuncture group were treated first with scalp acupuncture for 20 min, followed by general anesthesia, with scalp acupuncture lasted till the end of operation. The general anesthesia group were treated only with simple intratracheal anesthesia. Effects of scalp acupuncture on malondialdehyde (MDA) level and superoxide dismutase (SOD) activity at the operation stage were observed. RESULTS: After operation, MDA levels decreased significantly (P < 0.05) in the two groups, but with no difference between the two groups in the different values of MDA before and after treatment (P > 0.05); there was no significant change in SOD before and after treatment in the scalp acupuncture group, and significantly decreased in the general anesthesia group (P < 0.05). There was a significant difference between the two groups in the different value of SOD before and after the operation (P < 0.05). CONCLUSION: Scalp acupuncture can inhibit the decrease of SOD activity, reducing oxidative stress reaction at operation stage.

[Role of acupuncture anesthesia in operation of rectal cancer].

OBJECTIVE: To observe analgesic effect of acupuncture anesthesia. METHODS: Sixty-nine cases undergoing rectal cancer surgery were randomly divided into 3 groups, group I, II and III, 23 cases in each group. Both Zusanli (ST 36) and Sanyinjiao (SP 6) were selected for acupuncture anesthesia. Group I received general anesthesia after acupuncture induction, group II received acupuncture after general anesthesia, and group II received only general anesthesia. Minimum alveolar concentration (MAC) before and after operation was recorded. RESULTS: MAC was (1.35 +/- 0.19) vol% in the group I, (1.49 +/- 0.22) vol% in the group II and (1.64 +/- 0.27) vol% in the group III. Acupuncture before and after general anesthesia could decrease respectively by about 0.29% and 0.15% of MAC in rectal cancer surgery undergoing general anesthesia, with a very significant difference (P < 0.01) or a significant difference (P < 0.05) among the 3 groups. CONCLUSION: Acupuncture anesthesia has a certain adjuvant action for anesthesia and analgesia, and acupuncture before general anesthesia has a better action.

[Regeneration of autologous tissue-engineered cartilage by using basic-fibroblast growth factor in vitro culture].

OBJECTIVE: To investigate the effect of the basic fibroblast growth factor (b-FGF) to regenerate an autologous tissue-engineered cartilage in vitro. METHODS: The Cells were harvested from the elastic auricular cartilage of swine,and were plated at the concentration of 1 x 10(4) cells/cm2 , studied in vitro at two different media enviroments: Group I contained Ham's F-12 with supplements and b-FGF, Group II contained Ham's F-12 only with supplements. The passage 2 cells (after 12.75 +/- 1.26 days) were harvested and mixed with 30% pluronic F-127/Ham's F-12 at the concentration of 50 x 10(6) cells/ml. It was injected subcutaneously at 0.5 ml per implant. The implants were harvested 8 weeks after the vivo culture and examined with the histological stains. RESULTS: The chondrocytes displayed morphologically similar to the fibroblasts in the media containing basic-FGF. The number of cell doublings (after 12.75 +/- 1.26 days) in vitro culture was as the following: Group I, 70; Group II, 5.4. Eight 8 weeks after the vivo autologous implantation, the average weight (g) and volume (cm3) in each group was as the following: Group I, 0.371 g/0.370 cm3 Group II, 0.179 g/0.173 cm3 (P < 0.01). With the b-FGF in vitro culture, the cells were expanded by 70 times after 2 weeks. Histologically, all of the engineered cartilage in the two groups were similar to the native elastic cartilage. CONCLUSION: These results indicate that the basic-FGF could be used positively to enhance the quality and quantity of the seeding cells for the generation of the well-engineered cartilage.

A phase III study of adjuvant chemotherapy in advanced nasopharyngeal carcinoma patients.

PURPOSE: To evaluate the role of adjuvant chemotherapy in locally advanced nasopharyngeal carcinoma (NPC) patients, we conducted a randomized Phase III trial comparing radiotherapy (RT) followed by adjuvant chemotherapy to RT alone in patients with advanced NPC. METHODS AND MATERIALS: Between November 1994 and March 1999, 157 patients with Stage IV, M(0) (UICC/AJCC, 1992) advanced NPC disease were randomized to receive standard radiotherapy, as follows: 35-40 fractions, 1.8-2.0 Gy/fraction/day, 5 days/week, to a total dose 70-72 Gy with or without 9 weekly cycles of 24-h infusional chemotherapy (20 mg/m(2) cisplatin, 2,200 mg/m(2) 5-fluorouracil, and 120 mg/m(2) leucovorin) after RT. Of 157 patients enrolled, 154 (77 radiotherapy, 77 combined therapy) were evaluable for survival and toxicity analysis. RESULTS: With a median follow-up of 49.5 months, the 5-year overall survival and relapse-free survival rates were 60.5% vs. 54.5% (p = 0.5) and 49.5% vs. 54.4% (p = 0.38) for the radiotherapy-alone group and the combined radiotherapy and adjuvant chemotherapy group, respectively. The Cox regression showed that the hazard rates ratio of combined treatment to RT alone was 0.673 (p value = 0.232); the 95% confidence interval was 0.352 and 1.288, respectively. Patients who received combined treatment had a lower systemic relapse rate than radiotherapy-alone patients, according to relapse pattern analysis. The incidence of leukopenia (>or= Grade 3) occurred in 17 out of 819 (2.1%) cycles of weekly chemotherapy. No patient developed moderate to severe mucositis (>or= Grade 3). CONCLUSIONS: We conclude that adjuvant chemotherapy after RT for patients with advanced NPC has no benefit for overall survival or relapse-free survival.