TCR repertoire profiling of tumors, adjacent normal tissues, and peripheral blood predicts survival in nasopharyngeal carcinoma.

The T-cell immune responses in nasopharyngeal carcinoma patients have been extensively investigated recently for designing adoptive immunotherapy or immune checkpoint blockade therapy. However, the distribution characteristics of T cells associated with NPC pathogenesis are largely unknown. We performed deep sequencing for TCR repertoire profiling on matched tumor/adjacent normal tissue from 15 NPC patients and peripheral blood from 39 NPC patients, 39 patients with other nasopharyngeal diseases, and 33 healthy controls. We found that a lower diversity of TCR repertoire in tumors than paired tissues or a low similarity between the paired tissues was associated with a poor prognosis in NPC. A more diverse TCR repertoire was identified in the peripheral blood of NPC patients relative to the controls; this was related to a significant decrease in the proportion of high-frequency TCR clones in NPC. Higher diversity in peripheral blood of NPC patients was associated with a worse prognosis. Due to the peculiarity of the Vß gene usage patterns in the peripheral blood of NPC patients, 15 Vß genes were selected to distinguish NPC patients from controls by the least absolute shrinkage and selection operator analysis. We identified 11 clonotypes shared by tumors and peripheral blood samples from different NPC patients, defined as "NPC-associated" that might have value in adoptive immunotherapy. In conclusion, we here report the systematic and overall characteristics of the TCR repertoire in tumors, adjacent normal tissues, and peripheral blood of NPC patients. The data obtained may be relevant to future clinical studies in the setting of immunotherapy for NPC patients.

Changes of plasma cytokines and chemokines expression level in nasopharyngeal carcinoma patients after treatment with definitive intensity-modulated radiotherapy (IMRT).

BACKGROUND: Potential clinical application values of certain cytokines and chemokines that participate in the process of tumor growth, invasion, and metastasis have been reported. However, there still lack of biomarkers for a great many of malignancy. This study identified cytokines or chemokines involved in the occurrence and development of nasopharyngeal carcinoma (NPC), which might be a biomarker for noninvasive early diagnosis. METHODS: The plasma levels of 19 cytokines and chemokines were detected by the luminex liquid array-based multiplexed immunoassays in 39 NPC patients before and after treatment by definitive intensity-modulated radiotherapy (IMRT). RESULTS: Plasma levels of almost all of the 19 cytokines and chemokines in NPC patients were higher than healthy controls, while only IFN-γ, IL-1b IL-6, MCP-1, TNF-α, FKN, IL-12P70, IL-2, IL-5 and IP-10 showed significant differences. However, expression levels of most of the 19 cytokines and chemokines decreased after therapy, especially IFN-γ, IL-10, IL-1b, IL-6, IL-8, MCP-1, TNF-α, VEGF, IL-17A, IL-2, IL-5 and MIP-1b, have a dramatic decline. Taking together, plasma levels of IFN-γ, IL-1b, IL-6, MCP-1, TNF-α, IL-2 and IL-5 are significantly increased in NPC patients and dramatically decreased after treatment, suggesting these cytokines and chemokines might play important roles in the progress of NPC. More interestingly, the expression level of MPC-1 is significantly associated with clinical stage. CONCLUSION: MCP-1 might involve in the genesis and development process of NPC, which might serve as a noninvasive biomarker for early diagnosis.

Pretreatment Diffusion-Weighted MRI Can Predict the Response to Neoadjuvant Chemotherapy in Patients with Nasopharyngeal Carcinoma.

PURPOSE: To explore the potential of diffusion-weighted (DW) magnetic resonance imaging (MRI) using apparent diffusion coefficient (ADC) for predicting the response to neoadjuvant chemotherapy in nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: Ninety-two consecutive patients with NPC who underwent three cycles of neoadjuvant chemotherapy were retrospectively analyzed. DW and anatomical MRI were performed before and after neoadjuvant chemotherapy prior to radiotherapy. Pretreatment ADCs and percentage increases in ADC after chemotherapy were calculated for the primary lesions and metastatic adenopathies. Receiver operating characteristic curve analysis was used to select optimal pretreatment ADCs. RESULTS: Pretreatment mean ADCs were significantly lower for responders than for nonresponders (primary lesions, P = 0.012; metastatic adenopathies, P = 0.013). Mean percentage increases in ADC were higher for responders than for nonresponders (primary lesions, P = 0.008; metastatic adenopathies, P < 0.001). The optimal pretreatment primary lesion and metastatic adenopathy ADCs for differentiating responders from nonresponders were 0.897 × 10(-3) mm(2)/sec and 1.031 × 10(-3) mm(2)/sec, respectively. CONCLUSIONS: NPC patients with low pretreatment ADCs tend to respond better to neoadjuvant chemotherapy. Pretreatment ADCs could be used as a new pretreatment imaging biomarker of response to neoadjuvant chemotherapy.

Prognostic value of classifying parapharyngeal extension in nasopharyngeal carcinoma based on magnetic resonance imaging.

PURPOSE: To subclassify parapharyngeal extension in nasopharyngeal carcinoma (NPC) and investigate its prognostic value and staging categories based on magnetic resonance imaging (MRI). METHODS AND MATERIALS: Data from 1504 consecutive NPC patients treated with definitive-intent radiotherapy were analyzed retrospectively. Sites of parapharyngeal extension were defined by MRI. Overall survival (OS), local relapse-free survival (LRFS), and distant metastasis-free survival (DMFS) were calculated by the Kaplan-Meier method and compared with the log-rank test. Hazard consistency and hazard discrimination were determined by multivariate analysis with Cox proportional hazards models. RESULTS: 1104 patients (73.4%) had parapharyngeal extension; 1.7-63.8% had involvement of various anatomic sites. The hazard ratio for death was significantly higher with extensive parapharyngeal extension (lateral pterygoid muscle of masticator space and beyond or parotid space) than with mild extension (medial pterygoid muscle of masticator space, or carotid, prestyloid, and prevertebral or retropharyngeal space). OS, LRFS, and DMFS with extensive parapharyngeal extension were similar to those in T4 disease; OS, LRFS, and DMFS with mild parapharyngeal extension were significantly higher than in those T3 disease (all P ≤ 0.015). CONCLUSIONS: Parapharyngeal extension in NPC should be subclassified as mild or extensive, which should be regarded as stages T2 and T4 diseases, respectively.

MicroRNA-9 suppresses the sensitivity of CNE2 cells to ultraviolet radiation.

MicroRNA (miR)-9 has been demonstrated to regulate the radiosensitivity of tumor cells. In the present study, the mechanism by which miR-9 modulates the sensitivity of nasopharyngeal carcinoma (NPC) cells to ultraviolet (UV) radiation was investigated. The results demonstrated that exposure of NPC cells to UV light resulted in a significant increase in the expression of miR-9, and that CNE2 cells overexpressing miR-9 exhibited reduced levels of DNA damage and increased levels of total glutathione upon UV exposure. Accordingly, the inhibition of the expression of miR-9 promoted UV-induced DNA damage and apoptosis. Although miR-9 inhibited the expression of E-cadherin in the CNE2 cells and increased their resistance to UV radiation, the use of small interfering RNA to inhibit the expression of E-cadherin was not sufficient to decrease the radiosensitivity of the NPC cells. These data demonstrated that miR-9 did not modulate the sensitivity of the CNE2 cells to UV radiation through E-cadherin, but suggested that miR-9 regulated radiosensitivity through its effects on glutathione. These findings suggest that miR-9 may be a potential target for modulating the radiosensitivity of NPC cells.

Functionally distinct subsets of CD4⁺ regulatory T cells in patients with laryngeal squamous cell carcinoma are indicative of immune deregulation and disease progression.

CD4+ regulatory T cells (Tregs) mediate immune tolerance in laryngeal squamous cell carcinoma (LSCC). However, Tregs are functionally heterogeneous. Recently, we reported that three distinct Treg subsets (resting Tregs, activated Tregs and cytokine-secreting CD45RA-Foxp3lowCD4+ T cells) vary in the peripheral circulation of patients with head and neck squamous cell carcinoma (HNSCC); however, the potential implication of these Treg subsets in LSCC immunity is unclear. Here, we report that activated Tregs and cytokine­secreting CD45RA-Foxp3lowCD4+ T cells were increased in LSCC patients compared with healthy donors (HD) (p<0.001, p<0.001), whereas resting Tregs were decreased (p<0.001). Activated Tregs inhibited the proliferation of CD4+CD25- T cells (p<0.001) and secreted lower levels of interleukin-2 (p<0.001), interferon-γ (p<0.001) and tumor necrosis factor-α (p<0.001) compared with the cytokine-secreting CD45RA-Foxp3lowCD4+ T cells. Importantly, activated Treg prevalence was correlated with tumor stage (p=0.001) and nodal status (p=0.007). The prevalence of naïve CD4+ (p<0.001), naïve CD8+ (p=0.002), and Th1 T-cell subsets (p<0.001, p<0.001) was decreased in the LSCC patients. In conclusion, our findings showed that activated Tregs with suppressive activity are a distinct subset of Tregs in LSCC, and correlate with disease progression. Several immune system abnormalities in LSCC patients are represented by expansion of functionally activated Tregs, both in the circulation and tumor microenvironment along with decreased frequencies of naïve T-cell populations and Th1-cell populations.

Prognostic value of grading masticator space involvement in nasopharyngeal carcinoma according to MR imaging findings.

PURPOSE: To derive a suitable method for grading masticator space invasion in nasopharyngeal carcinoma on the basis of magnetic resonance (MR) images and to determine its prognostic value in patients undergoing intensity-modulated radiation therapy. MATERIALS AND METHODS: After institutional review board approval and informed consent were acquired, 808 patients with nasopharyngeal carcinoma who were treated with definitive intensity-modulated radiation therapy were analyzed retrospectively. The anatomic sites of masticator space involvement were identified with MR imaging. Overall survival, local relapse-free survival, and distant metastasis-free survival were calculated by using the Kaplan-Meier method and were compared by using the log-rank test. Potential prognostic factors were identified by means of multivariate analysis. RESULTS: Masticator space involvement was diagnosed in 163 of 808 patients (20.2%). Patients with lateral invasion (involvement of the lateral pterygoid muscle of the masticator space and beyond) had significantly poorer overall survival and distant metastasis-free survival than those with medial invasion (involvement of the medial pterygoid muscle of the masticator space) (P = .035 and P = .026, respectively). Furthermore, their overall survival, local relapse-free survival, and distant metastasis-free survival were significantly poorer compared with patients with stage T2 or T3 disease (all P ≤ .023) but similar to patients with stage T4 disease. The grade of masticator space involvement was an independent prognostic factor for overall survival, local relapse-free survival, and distant metastasis-free survival (all P ≤ .023). CONCLUSION: Masticator space involvement in nasopharyngeal carcinoma should be graded as medial (stage T2 disease) or lateral (stage T4 disease). This can facilitate staging of nasopharyngeal carcinoma and may be a suitable prognostic indicator of survival.

[Therapeutic effects of small fenestra stapedotomy with semiconductor diode laser: a comparison with microdrill].

OBJECTIVE: To compare the effects of small fenestra stapedotomy with semiconductor diode laser and microdrill in patients with otosclerosis. METHODS: Twenty-six patients (29 ears) undergoing stapedotomy with semiconductor diode laser and 19 patients (21 ears) with microdrill were compared for the hearing results and complication rates. RESULTS: No statistically significant differences were found in postoperative speech frequency and high frequency pure tone average in closing the air-bone gap between the two groups. The ears treated by stapedotomy with semiconductor diode laser showed significantly better preoperative minus the postoperative air-bone gap and milder dizziness. CONCLUSION: In spite of the good hearing outcomes in both groups, small fenestra stapedotomy with semiconductor diodelaser can achieve better results and reduce the incidence of complications.

[Perioperative management of severe obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To investigate the perioperative management of severe obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: Fifty-three patients with severe OSAHS received uvulopalatopharyngoplasty with uvula preservation and radiofrequency tongue base reduction. All the patients were treated with automated continuous positive airway pressure (CPAP) for 3-7 days before operation and automated antibiotic therapy administered in the oropharynx, with 24 h ECG monitoring postoperatively. Polysomnography were carried out before and 6 months after surgery. RESULTS: The preoperative apnea hypopnea index (AHI) and lowest SaO(2) (LSaO(2)) were 58.4-/+5.1/h and 0.650-/+0.059, respectively, which were 15.5-/+3.2/h and 0.864-/+0.064 at 6 months after surgery, respectively, showing significant changes after surgery (P<0.01). Dyspnea occurred in 2 cases after operation, intraoperative bleeding in 1 case, primary bleeding in 2 cases and hypertension crisis in 1 case. CONCLUSION: Severe OSAHS patients are subject to great surgical risk. Application of auto-CPAP before operation can significantly improve the patients' tolerance of surgery and anesthesia, and reduce the surgical risks and preoperative complications.

[Clinical application of uvulopalatopharyngoplasty with uvula preservation and tongue base radiofrequency reduction].

OBJECTIVE: To explore the effect of uvulopalatopharyngoplasty with uvula preservation and radiofrequency tongue base reduction for obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: Forty six patients with OSAHS were enrolled. One group (n = 22) of patients only received uvulopalatopharyngoplasty with uvula preservation, while the other group (n = 24) had both uvulopalatopharyngoplasty with uvula preservation and radiofrequency tongue base reduction. Polysomnography and distance between anterior pillars (DBAP), distance between posterior pillars (DBPP), length of roft palate, distance between tongue base and posterior pharyngeal wall (DBTP) were measured before and 6 months after surgery. RESULTS: The pre-operation apnea hypopnea index (AHI), x +/- s, lowest SaO2 (LSaO2) of first group were (56. 5 +/- 6. 0)/h, and 0.626 +/- 0.060 respectively, and 6 months after surgery, AHI was (23.7 +/- 2.7)/h, LSaO2 was 0.797 +/- 0.053. The pre-operation AHI, LSaO2 of second group were (58.4 +/- 5.1)/h, and 0.650 +/- 0.057 respectively, and 6 months after surgery, AHI was (15.5 +/- 3.2)/h, LSaO2 was 0.864 +/- 0.064. After surgery AHI and LSaO2 have changed in both groups (P<0.001). Six months after operation, DBAP and DBPP became withy, length of soft palate became short (P<0. 001). In one group the validity ratio is 72.7% (16/22), the other group the validity ratio is 87.5% (21/24) (P< 0.05), and pharyngeal posterior airway width (PPAW) became withy (P <0.001). CONCLUSIONS: For OSAHS patients, the obstructive regions should be evaluated. The combined surgery of uvulopalatopharyngoplasty with uvula preservation and radiofrequency tongue base reduction could have a better result.

[An experimental study of mucociliary clearance of trachea reconstructed with free jejunum].

OBJECTIVE: To investigate tracheal mucociliary transport change after reconstructed with free jejunum. METHODS: Twelve canine models of extensive circumferential tracheal defects reconstructed with revascularized jejuno combined with Ni-Ti alloy mesh tube were established. Every canine model was marked in cervical skin projecting the lower resected margin of trachea lumen and was injected 50% barium sulfate mucilage as a tracer into the trachea lumen under bronchoscopy. Record the time from tracer injected into trachea lumen to its arriving glottis (mucociliary transit time, MTT) and the length from tracer injected into trachea lumen to glottis (mucociliary transport length, MTL). Mucociliary transit rate (MTR), as MTL/ MTT, was calculated. The same procedure was performed at preoperative and postoperative 7th day, 1 month, 3 months and 6 months. RESULTS: There were statistical significance between preoperative MTR and 1 month postoperative MTR (P < 0.05). There were no differences between preoperative MTR and postoperative MTR at the 7th day, 1 month, 3 months and 6 months. There were also no differences between postoperative MTR at the 7th day and 1 month, 3 months and 6 months. CONCLUSION: In new tracheal tract reconstructed with free jejunum, MTR becomes normal at 3 months postoperatively.

[Salvage surgical approach for patients with post-radiation local recurrent nasopharyngeal carcinoma].

OBJECTIVE: To evaluate salvage surgical approaches and efficacy for post-radiation local recurrent nasopharyngeal carcinoma (NPC). METHODS: Thirteen patients with post-radiation local recurrent NPC underwent salvage surgical treatment by routes as transpalatal approach, nasal medial swing approach, maxillary swing approach and infratemporal fossa approach. All cases were followed up for 2 to 5 years. Analysis was done on the indications and efficacy of these 4 different approaching routes. RESULTS: No immediate operative complications occurred for all these 13 cases. Four patients with T1 and T2a operated via transpalatal approach and nasal medial swing approach survived more than 3 years. Five patients with T2b and T3 operated via maxillary swing approach. Among them, two patients died at second and 24th month after operation, one survived with tumor and died at 13rd month after operation, two were alive free of tumor for 2 and 4 years after operation. Four patients with T4 operated via infratemporal fossa approach. Among them, three died in 1 year, one was alive free of tumor for 2 years. CONCLUSIONS: Surgical approaches were decided by a comprehensive consideration of recurrent tumor site and invasive range to achieve the best operative site exposure with minimal traumatic damage.

[Experimental reconstruction of the canine trachea with shape-memory titanium-nickel alloy stent coupled with free jejunal graft].

OBJECTIVE: To explore the reconstruction method of extensive circumferential tracheal defects longer than 6.0 cm and evaluate the influence on pulmonary function from jejunal secretion. METHODS: Jejunal secretion model without extraneous nerve were established. 10 mongrel dogs were randomly divided into two groups. In group A, the nude stent made by shape-memory titanium-nickel alloy (SMA stent) was placed in the interior of the intestinal lumen. In group B, the SMA stent with silicone membrane was placed in the interior of the intestinal lumen. The secretion and histological chance of these jejunal were observed regularly. The cervical tracheal segment (6.5 cm) was replaced by the intestinal graft. In group C (6 mongrel dogs), the nude stent made by SMA stent was placed in the interior of the intestinal lumen. In group D (6 mongrel dogs and 6 Beagle dogs), the SMA stent with silicone membrane was placed in the interior of the intestinal lumen, the nude "C-shaped" SMA stent was placed out of the intestinal lumen, and the silicone stent was removed the fourth week after operation. In group C and group D, endoscopic and histological examinations were performed between the first week and eighth month. RESULTS: The secretory peak of Jejunal secretion model without extraneous nerve ranged from the first day to seventh day after operation. The jejunal secretion reduced gradually from 7th days after operation. The jejunal secretion remained steady after postoperative two months. In group C, endoscopic examination showed heavy proliferation of granulation in the tracheo-intestinal anastomosis. 4 dogs died between seventh day to second month. In group D, one dog died from ileus third month after operation. The other all survived operation. Gentle pneumonia happened to some dogs during 1-2 months after operation by X-ray examination. No one died of pneumonia result from hypersecretion. CONCLUSIONS: Reconstruction of the canine trachea with SMA stent with silicone membrane placed in the interior of the intestinal lumen together with the nude "C-shaped" SMA stent placed out of the intestinal lumen achieve satisfactory effect, the reconstructed trachea remain unblocked and this method of tracheal reconstruction may be relatively perfect and be expected for clinical application in future. The jejunal secretion didn't have severe influence on pulmonary function of experimental canine and couldn't cause experimental canine death.

Factor analysis for bleeding in endoscopic sinus surgery and the countermeasures.

This study was designed to define the possible risk factors for bleeding during endoscopic sinus surgery on the basis of a statistical analysis of 635 cases. It was found that the volume of intra-operative bleeding was closely related to the following factors including times of operation, surgical approach, perioperative treatment and the surgical skills on the part of the surgeons.