Objective To observe the effect of hyaluronic acid on the proliferation of and tyrosinase activity in melanocytes.Methods Normal primary human melanocytes were isolated from infant foreskin tissue and cultured.Different concentrations(0 to 10 g/L)of hyaluronic acid wero added to the culture medium immediately or 8 hours after the inoculation of melanocytes.MTT assay was performed to detect the proliferation of melanocytes,and tyrosinase activity was determined to evaluate the effect of hyaluronic acid on the melanin synthesis by melanocytes.Results The proliferation level (absorbance at 490 am,A490)of melanocytes was 0.14±0.02,0.37±0.08,0.45±0.11,0.49±0.07,0.55±0.12,0.52±0.11,0.49±0.07,0.39±0.05,0.19±0.03 and 0.01 4-0.01 when treated with hyaluronic acid of 0,0.008,0.016,0.313,0.625,1.250,2.500,5.000,7.500 and 10.000 g/L,respectively.The hyaluronic acid of 0.08 to 5 g/L markedly accelerated the proliferation of melanocytes,while that of 10 g/L inhibited their proliferation.The tyrosinase activity in melanocytes was promoted by hyaluronie acid of 0.2 to 5 g/L,but suppressed by that of 10 g/L.The proliferation of melanocytes treated with hyaluronic acid immediately after the inoculation was more rapid than that treated with hyaluronic acid 8 hours after the inoculation.Conclusion The hyaluronic acid of 0.2 to 5 g/L can enhance the proliferation of and tyrosinase activity in melanocytes.

Objective To evaluate the relationship between cell seeding density and clinical efficacy of autologous cultured melanocyte transplantation in the treatment of vitiligo.Methods A total of 632 patients with vitiligo were enrolled in this study,and randomly classified into 4 groups to be treated with transplantation of autologous cultured melanocytes at 4 different seeding densities respectively,i.e.,(3.0-4.9)× 104/cm2 (n =201),(5.0-7.9) × 104/cm2 (n =303),(8.0-9.9) × 104/m2 (n =82),(10.0-12.0) × 104/cm2 (n =46).Epidermal sheets were obtained by suction blister biopsy from the normal skin of the vitiligo patients,and subjected to the isolation and culture of melanocytes.After 2 to 5 passages,the cultured autologous melanocytes were transplanted at different seeding densities to vitiligous lesions,which were abraded previously by ultra-pulsed CO2 laser,of these patients.All the patients were followed for 6-12 months.Results At 6 months after the transplantation,52.85％of these patients achieved more than 90％ repigmentation,and 82.28％ more than 50％ repigmentation,with no differences in the cure rate and response rate between the 4 groups (both P ＜ 0.05).The percentage of patients obtaining excellent color matching was significantly higher in the group treated with transplantation of melanocytes at a seeding density of (5.0-7.9) × 104/cm2 than in the other 3 groups at 6,12 and 24 months after treatment (all P ＜ 0.05),and higher in all the 4 groups at 12-and 24-month points compared with the 6-month point (all P ＜ 0.05),but no statistical difference was observed between the 12-and 24-month point in any of these groups (all P ＞ 0.05).Conclusions The transplantation of autologous cultured pure melanocytes is effective for the treatment of stable vitiligo with the optimal cell seeding density of melanocytes being (5.0-7.9) × 104/cm2,and the color matching appears to improve with time.

Objective To retrospectively analyze the treatment outcomes and failure patterns in patients with head?and?neck cancer of unknown primary head?and?neck cancer of unknown primary, and to compare the efficacy between elective mucosal irradiation and ipsilateral neck treatment. Methods The clinical data of patients with head?and?neck cancer of unknown primary who were admitted from January 2007 to December 2013 were retrospectively collected. Thirty?one patients received elective pharyngeal mucosal irradiation and 61 patients only received ipsilateral neck treatment. The SPSS 19. 0 software was used for comparison of the survival and local control between the two groups. Results In the 92 patients, the median age was 57 years;79. 3% had metastasis to level Ⅱ lymph nodes;the median follow?up time was 36. 5 months;the 3?year overall survival, mucosal control, and neck control rates were 89. 0%, 86. 6%, and 82. 4%, respectively. Primary sites were found in 15 patients, containing nasopharynx in 4 patients, oropharynx in 3 patients, oral cavity in 3 patients, throat and hypopharyngeal part in 3 patients, maxillary sinus in 1 patient, and esophagus in 1 patient. The patients undergoing elective pharyngeal mucosal irradiation had significantly higher 3?year mucosal control and neck control rates than those undergoing ipsilateral neck treatment ( 100% vs. 74. 9%, P= 0. 040;87. 5% vs. 62. 2%, P= 0. 037 ) . There was no difference in the 3?year overall survival rate between the two groups ( 83. 5% vs. 88. 7%, P= 0. 910 ) . Conclusions For patients with head?and?neck cancer of unknown primary, elective pharyngeal mucosal irradiation can reduce the incidence of primary site and increase the neck control rate. A new standard for target volume delineation should be established as soon as possible for elective prophylactic pharyngeal mucosal irradiation.

Objective To evaluate the therapeutic effect of transplantation of autologous melanocytes cultured with individualized medium in vitiligo. Methods Donor skin was obtained by suction blisters from a normally pigmented area of the abdomen of 155 patients with vitiligo. The roof of the blisters was clipped and digested with trypsin, then the suspension of epidermal cells and melanocytes were cultured in Hu16 medium.The cell division time (DOT) and melanin content of cultured melanocytes were measured followed by the adjustment of concentration of fetal calf serum, cytokines and cAMP elevating agents based on the DOT,melanin content and morphology of melanocytes for the individualized culture of melanocytes. After 2 - 5 passages, melanocytes were harvested and inoculated into ultrapluse CO2 laser-denuded lesions. All patients were followed up for at least 6 months. Results One hundred and fifty-five vitiligo patients with 204 lesions were treated with transplantation of autologous melanocytes. Of the 155 patients, 119 received 1 session of transplantation, 36 received 2 to 4 session of transplantation. Cells were expanded by 50 - 80 times in vitro after individualized culture. Repigmentation was more than 50% in 84.8% of these lesions, more than 90% in 52.94% of the lesions. A homogeneous skin color was obtained in repigmented skin, and no scarring or other side effects were observed. No influence was noted on the outcome of transplantation for sex, age, course of disease or lesion size of patients. Segmental vitiligo showed better response than vitiligo vulgaris: the effective rate and cure rate were 93.62% and 65.96% respectively for segmental vitiligo, 82.16% and 49.04% respectively for vitiligo vulgaris. Lesions located on the arms and legs (not including elbows and knees) showed the best response, with a cure rate of 73.08%, whereas acral sites were the most difficult area to repigment, with a cure rate of just 25.93%. Conclusions Individualized culture can significantly increase the success rate of melanocyte culture and expanding times of melanocytes. Transplantation of cultured autologous melanocytes is an effective modality deserving clinical application in the treatment of stable vitiligo, with the advantage of treating large depigmented area with melanocytes from a small donor site.

Objective To investigate the relationship between nutritional status and quality of life ( QOL) in patients with nasopharyngeal carcinoma undergoing concurrent chemoradiotherapy, and to identify the optimal indices of body composition for evaluation of malnutrition. Methods Based on the European Society Parenteral and Enteral Nutrition ( ESPEN) consensus on the diagnosis of malnutrition and patient?generated subjective global assessment (PG?SGA), a prospective study was performed in 48 patients with nasopharyngeal carcinoma who received concurrent chemoradiotherapy in our hospital from 2014 to 2015. Changes in body composition and nutritional status were evaluated in those patients. The relationship between nutritional status and QOL was studied. The predictive factors for nutritional status in patients with nasopharyngeal carcinoma were explored. The correlation between datasets was evaluated by the Pearson correlation coefficient. The influencing factors for nutritional status were analyzed by Logistic regression. Results During concurrent chemoradiotherapy, some indices of body composition, including weight, body mass index ( BMI) , fat mass index ( FMI) , fat?free mass index ( FFMI) , body cell mass ( BCM) , skeletal muscle mass ( SM) , and phase angle ( PA) had different degrees of reduction ( P=0?00) , while the PG?SGA score gradually increased. The incidence of malnutrition in patients with nasopharyngeal carcinoma during radiotherapy was 2?1%?39?6% based on 2015 ESPEN consensus on the diagnosis of malnutrition and 12?5%?41?7% based on PG?SGA. These two methods showed good agreement at the fourth and sixth weeks of radiotherapy ( Kappa=0?911;Kappa=0?957) . The changes in QOL score were correlated with changes in FFMI and weight during radiotherapy ( r= 0?805, P= 0?00;r= 0?777, P= 0?00 ) . Logistic regression indicated that age, FMI, and FFMI were influencing factors for nutritional status ( P= 0?035, 0?013, 0?043) . Conclusions Patients with nasopharyngeal carcinoma have substantial nutritional deterioration during chemoradiotherapy. The nutritional status is closely associated with QOL. A prediction model of nutritional status can provide a comprehensive and accurate judgment of nutritional status in patients.

OBJECTIVETo investigate the treatment outcome of loco-regionally advanced squamous cell carcinoma of the head and neck (LA-SCCHN) after been treated with multi-modality approach since 2005 in our hospital and to explore the prognostic factors for treatment outcomes.

METHODSClinical data of 125 postoperative LA-SCCHN patients treated in our department with radiotherapy/chemoradiotherapy from May 2005 to December 2011 were collected and reviewed in this study. The radiotherapy technique was intensity-modulated radiotherapy (IMRT) (93.6%) and a minority of patients received 3D-conformal radiotherapy (3D-CRT).

RESULTSUp to January 6th, 2013, 124 patients were followed up with a median follow-up duration of 25 months. The 3-year overall survival (OS), disease-free survival (DFS), loco-regional control (LRC), distant metastasis-free survival (DMFS) were 69.7%, 56.1%, 80.8%, and 73.1%, respectively. A total of 37 patients died during the follow-up period. Among the 43 patients presented with treatment failure, 13 patients had loco-regional relapse, 20 patients had distant metastasis and 10 patients presented with both loco-regional and distant relapses. Distant metastasis accounted for the predominant cause of death. Lung and mediastinal lymph nodes are the most common sites involved by distant metastasis.Univariate analysis indicated that patients who underwent non-radical surgery, with larger size of invaded lymph nodes, higher N stage (N2b and above) and vascular tumor embolism had a lower OS (P = 0.001, 0.000, 0.032, 0.007, respectively). Patients who underwent neck dissection only, or those with higher N stage (N2b and above) or higher TNM stage or vascular tumor thrombi had higher distant metastasis rates (P = 0.017, 0.002, 0.008, 0.001, respectively). The multivariate analysis showed that non-radical surgery was an independent prognostic factor for OS (P = 0.001), larger size of invaded lymph nodes was an independent prognostic factor for poorer LRC (P = 0.001); higher N stage (N2b and above) or T4 stage and vascular tumor thrombi were independent prognostic factors for poorer distant metastasis-free survival (P = 0.035, 0.008 and 0.050, respectively).

CONCLUSIONSOur results indicate that multi-modality treatment for LA-SCCHN has achieved better outcome than before. Distant metastasis has become the predominant pattern of failure as well as the primary cause of death instead of loco-regional relapse as a result of improved local control modality. More efforts should be made to decrease the rate of distant metastasis in the future.

Adult ; Aged ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; pathology ; secondary ; therapy ; Cetuximab ; Cisplatin ; therapeutic use ; Combined Modality Therapy ; Disease-Free Survival ; Female ; Fluorouracil ; therapeutic use ; Follow-Up Studies ; Head and Neck Neoplasms ; pathology ; secondary ; therapy ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Neoplastic Cells, Circulating ; Radiotherapy, Conformal ; Radiotherapy, Intensity-Modulated ; Survival Rate ; Taxoids ; therapeutic use