Aim Toanalyzethekeyqualityandestablishmeth-ods for essential quality control of human recombinant ASPP2 adenovirus.Methods TheviralstructuralgeneofE2Bandtar-get gene of ASPP2 were identified by PCR;The number of virus particles was measured by UV-SDS methods;Infectious titer was determined by TCID50 assay;Target protein of ASPP2 was ob-served by Western blot assay;The biological effects of recombi-nant adenovirus on liver cancer cells were evaluated by MTT as-say;A549 cells were used to check replication of the competent adenovirus(RCA)by the observation of the cytopathic effect. Results PCRanalysisofE2BandASPP2wasinconsistent with theoretical values;Particle numbers of virus were 5. 6 × 1012 VP/mL,infectious titer was 2 ×1011 IU/mL and specific activity was 3. 5%;ASPP2 protein expression could be detected when cells were infected with virus for 24 h;Growth inhibition of liver cancer cells could be found by adding recombinant ASPP 2 adenovirus;The level of RCA was less than 1 RCA/3. 0 ×1010 VP,in line with the standards of China Food and Drug Adminis-tration(SFDA).Conclusion Thequalitycontrolmethodswere established aiming at key characters of human recombinant AS-PP2 adenovirus,which may provide foundations for its quality standard and future applications.

AIM: To investigate the amount and patterns of expressing CD69 , IL-4 and IFN-? on TCRV?24 +NKT cells, and compare with that of CD3 +T cells from human peripheral blood in response to in vitro stimulation. METHODS: The whole blood was stained with three-color immunofluorescence directly or after cultured with PDB+ionomycin(Ion) for 6 h, then the mononuclear cells were separated by lysing red blood cells. The expression rates of CD69, IL-4 and IFN-? on TCRV?24 +NKT cells and CD3 +T cells were estimated by flow cytometer. RESULTS: As a proportion of mature T cells, the ratio of TCRV?24 + NKT cells to CD3 +T cells was about(1.34?0.42)%. The expression rates of CD69 on TCRV?24 + NKT cells and CD3 +T cells in response to PDB + Ion for 6 h were (96.71?1.33)% and (98.60?0.47)%, respectively, while the ratio were (11.47?2.86)% and (1.07?0.45)% in the unstimulated group, and there were significant difference between them. The expression rates of IL-4 and IFN-? on TCRV?24 +NKT cells stimulated with PDB+Ion for 6 h were (48.62?2.44)% and (46.65?8.91)%, respectively ,which were significantly higher than that of unstimulated group [(31.57?3.31)%, (13.45?6.29)%] and that of stimulated CD3 +T cells, though the expression rates on stimulated CD3 +T cells were significantly higher than that of unstimulated CD3 +T cells. CONCLUSIONS: There is small amount of NKT cells in adult human peripheral blood. The expression rates of IFN-? and IL-4 on these lymphocytes are higher than CD3 +T cells, suggesting that NKT cells are important immunomodulatory cells in special microvironments.

Objective To investigate the clinical typing of nasopharyngeal carcinoma in patients treated by intensity-modulated radiation therapy (IMRT).Methods A retrospective analysis was performed on 333 patients with nasopharyngeal carcinoma who were initially treated in our hospital from 2003 to 2006 ;they had no distant metastasis and received IMRT.These patients were divided into 4 clinical types according to their prognosis:type Ⅰ (without local-regional recurrence and without distant metastasis),type Ⅱ (with local-regional recurrence and without distant metastasis),type Ⅲ (without local-regional recurrence and with distant metastasis),and type Ⅳ (with local-regional recurrence and with distant metastasis).Results Of all the patients,70.0％ (233) were of type Ⅰ,12.9％ (43) of type Ⅱ,16.5％ (55) of type Ⅲ,and 0.6％ (2) of type Ⅳ.Of 57 patients with stage Ⅰ-Ⅱ nasophayngeal carcinoma,86％ (49) were of type Ⅰ,11％ (6) of type Ⅱ,4％ (2) of type Ⅲ,and 0％ (0) of type Ⅳ,and of 276 patients with stage Ⅲ-Ⅳ nasopharyngeal carcinoma,66.7％ (184) were of type Ⅰ,13.4％ (37) of type Ⅱ,19.2％ (53) of type Ⅲ,and 0.7％ (2) of type Ⅳ,with significant differences between the two patient groups (P =0.007).Of the 69 patients who received IMRT alone,80％ (55) were of type Ⅰ,12％ (8) of type Ⅱ,9％(6) of type Ⅲ,and 0％ (0) of type Ⅳ; of the 218 patients who received IMRT combined with neoadjuvant plus concurrent chemotherapy,68.8％ (150) were of type Ⅰ,13.8％ (30) of type Ⅱ,16.5％(36) of type Ⅲ,and 0.9％ (2) of type Ⅳ; of the 46 patients who received IMRT combined with neoadjuvant plus adjuvant chemotherapy,61％ (28) were of type Ⅰ,11％ (5) of type Ⅱ,28％ (13) of type Ⅲ,and 0％ (0) of type Ⅳ.Conclusions In patients with early and advanced nasopharyngeal carcinoma,type Ⅰ is the most common,and type Ⅳ is the least common;type Ⅱ is more frequent than type Ⅲ in early patients,while type Ⅲ is more frequent than type Ⅱ in advanced patients.The percentage of type Ⅰ patients increases,while that of type Ⅱ-Ⅳ patients decreases,as compared with the data of those treated by conventional radiotherapy.

Objective To retrospectively analyze the outcomes,adverse events,and prognostic factors in elderly patients initially diagnosed with nasopharyngeal carcinoma (NPC) who receive radiotherapy and chemotherapy.Methods From January 2004 to April 2007,202 patients (over 60 years of age) initially diagnosed with NPC received radical radiotherapy or chemotherapy.These patients included 165 males (81.7％) and 37 females (18.3％),with a median age of 65 years (range 60-86 years).Of all patients,1 had stage Ⅰ NPC,25 had stage Ⅱ NPC,120 had stage Ⅲ NPC,and 56 had stage Ⅳa/Ⅳb NPC,according to the AJCC staging system (6th edition).All patients received a radiation dose of ≥66 Gy,and 146 patients (72.3％) with locally advanced NPC also received chemotherapy.The Cox model was used for multivariate analysis.Results The follow-up rate was 100％.The 5-year local control rate,regional lymph node control rate,distant metastasis-free survival (DMFS) rate,disease-free survival rate,and overall survival (OS) rate were 93％,94％,82％,74％,and 67％,respectively.The multivariate analysis revealed that T stage,N stage,and age were prognostic factors for OS and DMFS (P =0.006,0.002,and 0.008 ;P =0.020,0.002,and 0.022).The severe acute toxicities included grade 4 radiation dermatitis (1 patient,0.5％),grade 4 radiation mucositis (2 patients,1.0％),and grade 4 bone marrow suppression (5 patients,2.5％).Conclusions The elderly NPC patients can tolerate radiotherapy and chemotherapy well.Multimodality therapy should be adopted for the patients aged 60-74 years who have advanced-stage NPC and good general conditions.

Objective To analyze the long-term efficacy,prognostic factors and radiation sequela of nasopharyngeal carcinoma(NPC)treated with radiotherapy at the end of last century.Methods From January 1995 to December 1998,1706 newly diagnosed NPC patients treated with radiotherapy were included in the retrospective clinical analysis.There were 1081 patients treated with radiotherapy alone,625 with two to three circles of chemotherapy(5-Fu and DDP)before radiotherapy,23 with thermotherapy during radiotherapy and 162 with braehytherapy by 192Ir after external beam radiation.Results The 5-year overall survival. local control survival and disease-free survival rates were 67.60%.84.20%and 64.22%.respectively.The 5-year survival of patients with stage Ⅰ,Ⅱ,Ⅲand Ⅳ(the Fuzhou Staging,1992)were 100%,75.93%,66.47%and 49.34%.respectively.Cox regression analysis showed that the TNM classification,radiotherapy discontinuance,chemotherapy,sex,age and anemia before radiotherapy were the significant factors of survival.Conclusions Our experience shows that the main factors for the long term survival of NPC patients after radiotherapy are early TNM stage,young age,female,non-anemia before radiotherapy, radiotherapy continuance and chemotherapy.

Objective To evaluate the dosimetric characteristics of hippocampal formation (HF) in the intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC).Methods Fifty-nine NPC patients underwent IMRT.Simultaneous integrated boost technology was used to determine the doses for the target areas.The dose ranges of the HF were collected by dose-volume histogram.The influence of T stage on the exposure doses ( Dmax,Dmean,D20,V10,V20,V30,and V40 ) were compared.Results The maximum dose for the HF (Dmax) ranged from 11.1 to 78.2 Gy(F =24.2,P ＜0.05) and the Dmean ranged from 3.2 to 44.6 Gy ( F =16.3,P ＜ 0.05 ).The Dmax and Dmean of the T1-2 stage patients were (40.8 ±9.4) Gy and ( 12.5 ±5.1 ) Gy,respectively,both significantly lower than those of the T3-4 stage patients [ (58.6± 14.8) Gy,(20.9± 9.3 ) Gy].The mean exposed volume of the T4 stage patients was significantly larger than that of the T1 and T2 stages patients.Conclusions In the IMRT of NPC,the HF receives rather high irradiation dose.T stage is the main factor influencing the dose,especially T3 and T4 stages deserve serious attention.

Objective To evaluate the efficacy and safety of actovegin against acute oral mucositis through a randomized controlled multicenter trial for nasopharyngeal carcinoma(NPC) patients treated by chemo-radiotherapy. Methods From February 2006 to May 2007,a total of 161 patients with newly diagnosed stage Ⅱ-ⅣA(1992 Fuzhou Stage) NPC were randomly assigned to the prevention group,the treatment group and the control group. All patients received current chemo-radiotherapy ± neoadjuvant chemotherapy. Radiation technique and dose were similar among the three groups. Intravenous infusion of aeovegin was started when radiation started in the prevention group and when grade 2 mueositis occurred in the treatment group,which was given 30 ml daily ,5 times per week until the end of radiotherapy. Criteria of NCI CTC 2.0 and VRS were used to evaluate acute oral mueositis and pain degree,respectively. Results 154 patients were eligible for the efficacy analysis,including 49 in the prevention group,53 in the treatment group and 52 in the control group. In the prevention group and the control group, the incidence was 31% and 56% (P=0.011) for grade 3-4 mucositis,59% and 83% (P=0.009) for grade 2-3 pain. In the treatment group and the control group,the corresponding number was 38% and 60% (P=0.023) ,70% and 90%, (P=0.014). The prevention group had a lower incidence(P=0.021) and longer average interval(P=0.009) of grade 2 mucositis when comparing with the control group. No drug-related adverse event was observed. Conclusions Prophylactic or therapeutic use of actovegin by intravenous infusion can significantly reduce the severity of ehemo-radiotherapy induced oral mucositis and pain. The prophylactic use may also postpone and decrease the incidence of grade 2 mucositis,which deserves clinic application.

Aim To isolate and purify a novel plasminogen activator(PA)from Gloydius brevicaudus venom(GBV)and study characterization and biological activities of GBV-PA.Methods Affinity chromatography in Benzamidine Sepharose 6B(AC)and Lichrospher C-18 4.6/250 reversed phase chromatography(RPC)were used for isolation and purification;SDS-PAGE was used to detect molecular weight(MW);Disc polyacrylamide gel eletrophoresis was used to measure the point of isoelectric(pI);Chromogenic substrate method was used to observe the biological activities.Results A novel GBV-PA which its purification reached the homogeneity level was isolated and purified from GBV by AC and RPC;The MW of the novel GBV-PA was 3.26?104 and the pI was 5.2;The novel GBV-PA activated human plasminogen specifically and the special activity was 2.87 t-PA IU?mg-1;Moreover,our results indicated that this novel GBV-PA was a serine proteinase which had no affinity to fibrin.Conclusion A novel GBV-PA that can be isolated and purificated from GBV by AC and RPC was proved to be a serine protease and has no affinity to fibrin.

Background and purpose:Nasopharyngeal carcinoma usually occurs in people of states of the Southern China. Chemoradiotherapy plays an important role in the therapy of advanced nasopharyngeal carcinoma. However, chemoradiotherapy causes more toxic side effects than radiation therapy alone, which affects the therapy. Now symptomatic treatment and nutrition supports are common ways in the clinic in order to improve the tolerance of patients for the therapy, but with little effect. To evaluate the clinical significance of percutaneous endoscopic gastrostomy (PEG) in advanced nasopharyngeal carcinoma. Methods: From Oct. 2010 to Jun. 2012, a total of 71 patients with advanced nasopharyngeal carcinoma who received PEG before chemoradiotherapy were enrolled. During chemoradiotherapy, gastrostomy diet and nursing were supplied, adverse events, tolerance and nutrition indicators including weight and alcohol of human albumin were detected as well. Results:PEG were performed successfully on 68 patients, but failed in the other 3 patients. Of the 68 patients, 4 have not received concurrent chemotherapy because of liver metastases and liver function damage, 51 of the remaining 64 patients could completely ifnished 3 cycles of concurrent chemotherapy, with the completion rate of concurrent chemotherapy at 79.69%. The weight change ranged from-11.86%to 0.83%, with a mean value of-5.32%±2.99%. Paired-sample t test of human albumin before and after the treatment showed no significant difference (P=0.742). Grade Ⅱ radioactive oral cavity mucositis appeared in 60.29%patients (41/68), and 22.06%patients (15/68) suffered gradeⅢradioactive oral cavity mucositis.Conclusion:For patients with advanced nasopharyngeal carcinoma, preventative PEG improved the tolerance of chemoradiotherapy, reduce the incidence of adverse events. The period of therapy interruption caused by sever adverse event were shortened as well, PEG also increased the completion rate of concurrent chemotherapy. Nutritional status and living quality of patients are improved. In addition, PEG is a safe, economic and simple method.

Objective To measure the set-up errors of patients with head and neck (H&N) cancer during the image guided intensity-modulated radiotherapy (IMRT) treatment and analyze the impact of setup errors on dose distribution ; then to further investigate the necessity of adjustment online for H&N cancer during IMRT treatment.Methods Cone-beam CT (CBCT) scanning of thirty patients with H&N cancer were acquired by once weekly with a total of 6 times during IMRT treatment.The CBCT images and the original planning CT images were matched by the bony structure and worked out the translational errors of the x,y,z axis,as well as rotational errors.The dose distributions were recalculated based on the data of each setup error.The dose of planning target volume (PTV) and organs at risk were calculated in the replanning,and than compared with the original plan by paired t-test.Results The mean value of x,y,z axis translational set-up errors were ( 1.06 ± 0.95 ) mm,( 0.95 ± 0.77 ) mm and ( 1.31 ± 1.07 ) mm,respectively.The rotational error of x,y,z axis were ( 1.04 ±0.791 ),( 1.06 ±0.89) and (0.81 ±0.61 ),respectively.PTV 95％ volume dose ( D95 ) and PTV minimal dose of replanning for 6 times set-up were lower than original plan (6526.6 cGy:6630.3 cGy,t =3.98,P =0.000 and 5632.6 cGy:5792.5 cGy,t =- 2.89,P =0.007).Brain stem received 45 Gydose volume ( V45 ) and 1％ brain stem volume dose ( D01 )were higher than original plan ( 3.54％:2.75％,t =3.84,P =0.001 and 5129.7 cGy:4919.3 cGy,t =4.36,P =0.000).Conclusions The set-up errors led to the dose of PTV D95 obviously insufficient and significantly increased V45,D01 of the brainstem.So,adjustment online is necessary for H&N cancer during IMRT treatment.