Objective To construct the recombinant adenovirus encoding human IL-24 gene for future gene therapy. Methods The human IL-24 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP. The target gene was detected by polymerase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. The expression of target protein was measured by the method of immunohistochemistry. Results Restriction endonuclease and PCR analysis confirmed that the human IL-24 gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.2?10 10 pfu/ml. The adenovirus has a strong effect on A549 cells and human IL-24 can express in it. Conclusion The recombinant adenovirus containing human IL-24 gene was successfully constructed by the method of homogenous recombination in bacteria.

Objective To construct the recombinant adenovirus encoding human p16 gene for future gene therapy.Methods The human p16 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria.Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP.The target gene was detected by polymerase chain reaction(PCR).The titer and its infection rate were determined using the green fluorescent protein(GFP) expression in the shuttle plasmid.Results Restriction endonuclease and PCR analysis confirmed that the human p16 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 6.1?10~(10)pfu/ml.The adenovirus has a strong effect on human fibroblast cells.Conclusion The recombinant adenovirus containing human p16 gene was successfully constructed by the method of homogenous recombination in bacteria.

Background and purpose:Adenoid cystic carcinoma is a rare tumor from salivary gland tissues; the incidence is about 1% to 2% of all head and neck malignant tumors. The incidence of adenoid cystic carcinoma of nasal sinuses is lower. Its characteristics include slow growth, less lymph node metastasis, easy to tissue invasion and growing along the nerves. It has high rate of recurrence and distant metastasis. Patients with the disease always have poor prognosis. This study aimed to investigate the prognosis and its impacting factors in Chinese patients with adenoid cystic carcinoma of nasal cavity and paranasal sinuses.Methods:This was a single center, prospective, observational study in Chinese patients with adenoid cystic carcinoma of nasal cavity and paranasal sinuses. Total 104 patients who received radiotherapy during the period between Sep. 2000 and Nov. 2012 were included and followed up for median 5.1 years. Kaplan-Meier, log-rank test, and COX proportional hazards model were used for survival-related analysis. Results:Mean age of patients was (54.5±11.5) years with equal numbers of males and females. The most common primary site was nasal cavity (63.5%) followed by maxillary sinus (29.8%). 76.0% of patients were at clinical stagesⅢ-Ⅳ, 56.7% of patients with positive surgical margin and 34.6% of patients with positive nerve invasion. In terms of treatment regimens, 67.3% of patients received postoperative radiotherapy, 27.9% received preoperative radiotherapy, only 4.8% received pure radiotherapy, and 29.8% received chemotherapy combined with surgery and radiotherapy.Conformal radiation therapy (CRT) was the most common radiotherapy which was used in 81.7% of patients, followed by intensity-modulated radiation therapy (IMRT) which was used in 18.3% of patients. Finally, 18 patients had recur-rence and 28 patients had distant metastasis. The most common metastatic site was lung, and nerve invasion was the independent risk factor for recurrence or metastasis (P=0.000 2). The overall survival rates of 5 and 10 years were 77% (95%CI: 68.7%-85.3%) and 67.8% (95%CI: 57.8%-77.9%), respectively. The disease-free survival rates of 5 years and 10 years were 57.8% (95%CI: 48.0%-67.7%) and 56.4% (95%CI: 46.3%-66.4%), respectively. Recurrence or distant metastasis was critical risk factor for overall survival (HR=60.1, 95%CI: 8.15-443.1,P<0.0001). Positive nerve invasion was a signiifcant factor for disease-free survival (HR=2.99, 95%CI: 1.642-5.445,P=0.0002). The primary sites, clinical stage, status of surgical margin, or radiotherapy methods had no impact on the prognosis of patients with adenoid cystic carcinoma of nasal cavity and paranasal sinuses.Conclusion:Positive nerve invasion, recurrence or distant metastasis may be important factors affecting the prognosis of Chinese patients with adenoid cystic carcinoma of nasal cavity and nasal sinuses.

ObjectiveTo evaluate the reliahility and validity of the chronic HBV-infections related stigma scale.MethodsThe initial items and construct of the scale were developed according to theoretical analysis and interviews of experts and patients.A total of 151 patients with chronic HBV-infection were administered by convenient sampling method in this pilot study. The reliability and the validity of the scale were then evaluated.ResultsThe response rate of the scale was 94.5％.The Cronbach α coefficients of all dimeusions ranged from 0.75-0.87.The results of correlation analysis showed that there were higher correlation coefficients ( r ranged from 0.62-0.86) between items and their hypothesized subscales than those with other subscales ( r ranged from 0.14-0.55).The scale distinguished between patients with low subscale scores ( the subscale scores were ( 1.89 ±0.30 ),( 1.86 ± 0.29 ),( 1.96 ± 0.23 ),( 2.29 ± 0.45 ),( 1.59 ± 0.42 ) independently) and those with high subscale scores(the subscalc scores were (3.62 ±0.44),(3.99 ±0.41 ),(3.79 ±0.37),(4.13 ±0.34),(3.10 ±0.53 ) independently) (P ＜ 0.01 ).Confirmatory factor analysis showed that the main indices of goodness of fit CFI was 0.94,NNFI 0.92,RMSEA 0.087.ConclusionThe chronic HBV-infections related stigma has good psychometric properties regarding to reliability and validity.

Objective To investigate the clinical features and survival status of mucosal melanoma of the nasal cavity and paranasal sinuses and to analyze the prognostic factors.Methods A retrospective analysis was performed on the clinical data of 94 patients with mucosal melanoma of the nasal cavity and paranasal sinuses treated from January 2000 to December 2012.Of the 94 patients,50 were male,and 44 were female.The median age of onset was 60 years (range,26-85 years).The primary sites were nasal cavity (86 patients),maxillary sinus (7 patients),and nasopharynx (1 patient).Cervical lymph node metastasis was observed in 10 patients (7 patients before treatment,2 patients during treatment,and 1 patient after treatment).No patient had distant metastasis.Patients were treated with surgery ± radiotherapy.The Kaplan-Meier method was used to calculate survival rates,and the logrank test was used for univariate prognostic analysis;the Cox regression model was used for multivariate prognostic analysis.Results The 1-,3-,and 5-year sample sizes were 80,54,and 50,respectively.The 1-,3-,and 5-year disease-related survival rates were 71％,33％,and 22％,respectively.Univariate analysis showed that the prognostic factors were age over 55 years (P =0.034),involvement of the posterior naris (P =0.011),involvement of the maxillary sinus (P =0.009),involvement of the hard palate (P =0.003),cervical lymph node metastasis (P =0.001),and therapeutic method (P =0.038).Multivariate analysis showed that the prognostic factors were involvement of the posterior naris (P =0.027),involvement of the orbit (P =0.005),and involvement of the hard palate (P =0.003).Conclusions The distant metastasis and local recurrence rates are high among patients with mucosal melanoma of the nasal cavity and paranasal sinuses,so combination therapy is imperative.Cervical lymph node metastasis rate is low.Rational clinical staging needs to be further explored.

OBJECTIVE: To investigate the distribution patterns of microvascular density(MVD) and lymph vessel density (LVD) in nasopharyngeal carcinoma, and to discuss their relationships with tumor recurrence-metastasis and prognosis. METHOD: One hundred and six patients with pathologically proved nasopharyngeal carcinoma were included in this study. Immunostainings for the vascular endothelial marker CD34 and the lymph vessel marker D2-40 were performed. The microvascular and lymph vessels within the tumor and the peritumoral area were respectively quantified with computer assisted morphometric analysis. RESULT: Both microvascular and lymph vessels were positive in the specimen of 106 nasopharyngeal carcinoma patients, with MVD and LVD higher in the peritumoral area than in the tumor. Univariate analysis showed that intra-tumor MVD,intra-tumor and peritumor LVD were correlated with nasopharyngeal carcinoma recurrence-metastasis (P < 0.05), while no significant correlation was found between tumor recurrence-metastasis and age, gender, T stage, lymph node stage, clinical stage and peritumor MVD (P > 0.05). Multivariate analysis demonstrated that intra-tumor MVD and peri-tumor LVD were two independent risk factors of nasopharyngeal carcinoma recurrence-metastasis. Higher intra-tumor MVD was associated with adverse prognosis of nasopharyngeal carcinoma (P < 0.05). CONCLUSION High MVD and LVD may be of significance in predicting poor prognosis in patients with nasopharyngeal carcinoma. Intra-tumor and peri-tumor microvessels may play different roles in the tumor recurrence and metastasis. Intra-tumor MVD and peri-tumor LVD should be the therapeutic targets in the nasopharyngeal carcinoma.
Adolescent ; Adult ; Aged ; Carcinoma, Squamous Cell ; blood supply ; pathology ; Child ; Female ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Nasopharyngeal Neoplasms ; blood supply ; pathology ; Neoplasm Staging ; Neovascularization, Pathologic ; Prognosis ; Young Adult

OBJECTIVE: To evaluate the therapeutic effect of malignant tumor of nasal cavity and sinus that based on modern radiotherapy. METHOD: Two hundred cases of malignant tumor of nasal cavity and sinus were retrospectively analyzed from 2004 to 2007. All the cases were location by CT simulator system and were treated with three dimensional conformal radiotherapy. RESULT: Among malignant tumor of nasal cavity and sinus, the incidence for locations was nasal cavity > maxillary sinus > ethmoid sinus > sphenoid sinus; the incidence for the type of pathology was squamous cell carcinoma > adenocarcinoma > olfactory neuroblastoma and olfactory esthesioneuroepithelioma > malignant melanoma > rhabdomyosarcoma; the incidence for general metastasis was rhabdomyosarcoma > malignant melanoma > adenocarcinoma, inverted papilloma and malignant changes > squamous cell carcinoma, olfactory neuroblastoma and olfactory esthesioneuroepithelioma. No severe radiation-related complication were found. CONCLUSION Clinical stage, pathological type were the important factors effecting the prognosis of patients with malignant tumor of nasal cavity and sinus. Three dimensional conformal radiotherapy based on CT simulator system could improve therapeutic effect and protect the normal tissue very well.
Adenocarcinoma ; pathology ; radiotherapy ; Adolescent ; Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; radiotherapy ; Esthesioneuroblastoma, Olfactory ; pathology ; radiotherapy ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; Nose Neoplasms ; pathology ; radiotherapy ; Paranasal Sinus Neoplasms ; pathology ; radiotherapy ; Prognosis ; Radiotherapy, Conformal ; methods ; Retrospective Studies ; Survival Rate ; Young Adult

Objective:To evaluate the survival prognosis for T 1 stage nasopharyngeal carcinoma patients complicated with different stages of cervical lymph node metastasis, aiming to provide reference for optimizing the treatment plan. Methods:Clinical data of 413 patients in non-keratinizing carcinoma and undifferentiated locally early nasopharyngeal carcinoma (T 1N 0-3M 0-1) undergoing radiotherapy alone or radiochemotherapy in Department of Radiation Oncology of our hospital from January 2014 to December 2019 were retrospectively analyzed. The survival analyses were performed with Kaplan-Meier method and statistically compared using the log-rank test. Results:Of all patients, 291 were male, and 122 were female (aged from 9 to 78 years old) with a median age of 51 years old. All patients were diagnosed with T 1N 0-3M 0-1 nasopharyngeal carcinoma. In the TNM stage grouping system, 48(11.6%) patients were classified as stage Ⅰ (T 1N 0M 0), 158(38.2%) cases of stage Ⅱ(T 1N 1M 0), 162(39.2%) cases of stage Ⅲ(T 1N 2M 0), and 45(10.9%) cases of stage Ⅳ A to Ⅳ B(T 1N 3M 0/T 1N xM 1). Eight patients (1.9%) with stage Ⅳ B had metastasis at presentation. The lymph node positivity rate of all patients reached up to 88.1%. Seven patients received three-dimensional conformal radiotherapy, 371 cases of intensity-modulated radiotherapy and 35 cases of volumetric-modulated arc therapy. The 5-year overall survival rate was (95.9±1.2)% and with 100% for T 1N 0M 0 patients, (99.2±0.8)% for T 1N 1M 0 patients, (95.1±2.2)% for T 1N 2M 0 patients and (87.9±6.6)% for T 1N 3M 0 patients, respectively. Primary distant metastasis and N 3 stage were significantly correlated with poor prognosis (both P<0.05). The most common long-term side effect of radiotherapy was xerostomia with an incidence rate of 18.6%(17.9% for grade 1 toxicity), followed by hearing damage and tooth discomfort. Only 2 patients developed Grade Ⅲ toxic reactions, manifested as complete hearing loss. Conclusions:Although T 1 nasopharyngeal carcinoma patients have a high propensity of cervical node metastasis, favorable clinical prognosis can be obtained after radiotherapy alone. Moreover, the long-term side effects under precision radiation exert no severe effect upon the quality of life of patients.

Objective: To study the efficacy, safety and long-term outcomes of integrated strategy of bortezomib-based induction regimens followed by autologous hematopoietic stem cell (ASCT) and maintenance therapy in Chinese multiple myeloma (MM) patients. Methods: 200 MM patients receiving integrated strategy of bortezomib--based induction regimens followed by ASCT and maintenance therapy were retrospectively and prospectively analyzed from December 1. 2006 to April 30. 2018. Results: The complete remission rates (CR) and better than very good partial remission rates (VGPR) after induction therapy, transplantation and maintenance therapy were respectively 31% and 75.5%, 51.8% and 87.7%,73.6% and 93.4%. There was no difference between 4 cycles and more than 5 cycles induction chemotherapy. The negative rate of MRD detection by flow cytometry was 17.6% and 38.2% respectively after induction and 3 months after transplantation. The negative rate of MRD gradually increased during the maintenance therapy. The success rate of high dose CTX combined with G-CSF mobilization was 95.5% and transplantation related mortality (TRM) was zero. The median time to progress (TTP) was 75.3 months and the median overall survival (OS) was 99.5 months. TTP of patients obtaining CR and negative MRD after induction were longer that those of no CR and positive MRD. TTP and OS of patients receiving triple-drug induction and ASCT in early stage were longer than those of double-drug induction and ASCT in late stage. LDH≥240 U/L, high risk cytogenetics, ISS II+III stage and HBsAg positive were prognostic factors at diagnosis. However, only MRD and high risk cytogenetics were independent prognostic factors after transplantation and maintenance therapy. The clinical characteristics of patients of TTP ≥6 years were listed below: light-chain type M protein, ISS I stage, normal level of hemoglobin and platelet, normal LDH, HBsAg negative, chromosome 17p-negative, good response and sustained good response. Conclusions Integrated strategy of bortezomib-based induction regimens followed by ASCT and maintenance therapy can significantly improve the short-term and long-term efficacy. The prognostic factors of TTP in different disease stages were different. Response to treatment, especially MRD, played a more important role in prognostic factors.