Objective To investigate the role of hydroxyapatite nanoparticle (nHAP) in the gene transfection of human colorectal cancer cell line SW480/M5 and the possible mechanisms.Methods The combination and protection of nHAP-Mg2+ to DNA were analyzed by gelose gelatin electrophoresis.Liposome and nHAP modified by magnesium chloride was combined,and the PEGFP-N1 plasmids were transfected into SW480/M5 cells.The gene transfection rate and the mean fluorescence intensity were observed by flow cytometry.The effect of nHAP-Mg2+ on the growth of the cells were studied by MTT.Results At appropriate proportion,nHAP-Mg2+ could combine the plasmids compeletly and protected the DNA.The gene could not be transferred by nHAP-Mg2+ alone.Combining the nanoparticles and liposome,the gene could be transferred very efficiently and the transfection rates were significantly higher than the liposome (P ＜ 0.05).The inhibition of cell growth was increased along with the concentration of nHAP-Mg2+ wether it was used alone or with the combination of liposome (P ＜ 0.05).Conclusions nHAP-Mg2+ has the ability to combining and protecting DNA and can be used to transfer gene as the adjunct carrier of liposome for the gene therapy of tumor cells to elevate the gene tansfection and expression rate and also enhance the anti-tumor effection.

Objective To investigate the pathological change in articular cartilages after firearm injury.Methods Four rabbits from 28 New Zealand healthy rabbits were chosen as control group and subjected to joint capsule incision only. Another 24 rabbits were equally divided into 6 experimental groups( groups B to G) and subjected to medial femoral condyle cartilage surface damage by the nail gun.After the operation, their specimens were collected after 6 h,3 d,7 d,14 d,28 d and 56 d, respectively.Tissue sections were observed and stained by HE staining and toluidine blue staining.The histolopathological changes in articular cartilage after firearm injury were detected.Results The color of articular cartilages in experimental groups became lighter, the cell number increased but then decreased, the articular cartilage layer disappeared, the cell shape became uneven, cells began to cluster and the Mankin score increased, and the statistical differences between experimental groups and control group were significant.Conclusion The histological pathological changes in articular cartilages after fiream injury seem to follow some pattern.The degeneration seems obvious after 7 days and then becomes heavier.

Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20:sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20: sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20:sTRAIL were secreted from MSC. scFvCD20:sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20:sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20:sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20:sTRAIL, pLenR. ISZ:sTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20:sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20:sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20:sTRAIL,and thus locally concentrated scFvCD20:sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20:sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .

AIM:To investigate whether oxidative stress is able to induce autophagy in mesenchymal stem cells (MSCs), and to explore the effects of autophagy on MSC proliferation and apoptosis under oxidative stress circumstance as well as the underlying mechanism for promoting the therapeutic effects of transplanted MSCs on treating diabetes mellitus e -rectile dysfunction ( DMED) .METHODS: Hydrogen peroxide ( H2 O2 ) was applied to simulate the oxidative stress cir-cumstance.The effects of H2 O2 at concentration of 0, 50, 100, 200, 400μmol/L on the viability of MSCs were tested by the method of Trypan blue exclusion and MTT assay respectively .The methods of MTT assay , Western blot and transmis-sion electron microscope ( TEM) were used to explore the effects of H 2 O2 on MSC apoptosis and autophagy .RESULTS:The proliferation of MSCs was obviously inhibited by H 2 O2 in a dose-dependent manner ( P<0.01) and the 50%inhibiting concentration (IC50) was (384.58 ±16.89) μmol/L.H2O2 induced apoptosis and autophay of MSCs .The proliferation rate of MSCs was suppressed by H 2 O2 significantly ( P<0.05 ) , with a further decline by blockade of autophagy ( P<0.05) whereas increased by blockade of apoptosis (P<0.05).H2O2 induced MSCs apoptosis obviously (P<0.05), with an augment of apoptosis ( P<0.05) by blockade of autophagy .Furthermore, the H2 O2 increased expression of cleaved caspase-3 and cleavage of poly ADP-ribose polymerase 1 (PARP1), Which were decreased by apoptosis blockade whereas were enhanced by blockade of autopahgy .CONCLUSION:Oxidative stress plays a dual role in MSC survival , which in-duces MSC apoptosis and autophagy .Moreover , blockade of autophagy intensifies MSC apoptosis .Therefore , it is a promis-ing method to ameliorate the effects of stem-cell based therapy on DMED by enhancing protective autophagy to increase the survival rate of transplanted MSCs against oxidative stress circumstance caused by diabetes mellitus .

Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.

To elucidate the effects of Zearalenone(ZEA) on proliferation and cell cycle of cultured thymic epithelial cells in mice,trypan blue staining and flow cytometric analysis were performed.At the concentrations from 1 to 25 mg/L,ZEA displayed a significant inhibitory action to proliferation of thymic epithelial cells in its dose-and timedependent manner.Higher doses(10-25 rag/L)ZEA could induce a profound increase in G2/M phase with arrest of thymic epithelial cells in the G2/M phase in a dose-dependent manner.In conclusion,ZEA could be assumed that there were toxic effects on the thymie epithelial cells of mice in vitro.

Objective To study the effect of different dose fractionation on overall survival in patients with limited-stage small cell lung cancer (LS-SCLC). Methods LS-SCLC patients treated with radical combined chemotherapy and radiotherapy (RT) between January 2001 and Dec 2007 were analyzed retrospectively. According to the dose fractionation schemes, patients were divided into three groups:conventional fractionated RT (1. 8 -2.0 Gy,once daily), hyperfractionated RT (1.4 Gy, twice daily) and hypofractionated RT (2. 5 Gy,once daily). Overall survival, disease free survival and pattern of failures of the three groups were compared. A total of 177 patients were enrolled, including 63 patients in conventional fractionated RT group, 79 in hyperfractionated RT group and 35 in hypofractionated RT group. Results The overall follow-up rate was 96. 6%. The patient numbers with follow-up of more than 2 and 5 years were 153 and 92, respectively. The median survival time of the entire group was 22. 4 months, and the 2-and 5-year survival rates were 43.4% and 23. 5%, respectively. The 2-year survival rates for three groups were 31%, 46% and 59% (x2 =7.94,P=0.019), respectively. The 2-year disease free survival for three groups were 20%, 31% and 40% ( x2 = 4. 86, P = 0. 088 ), respectively. In the pairwise comparisons,patients in hypofractionated RT group have better survival than those in conventional fractionated RT group ( x2 = 7. 81, P = 0. 005 ), the effect of hyperfractionated RT group lies between the hypo-and the conventional fractionated RT groups, but no significant differences were detected ( x2 = 2. 31, P = 0. 128; x2 = 2. 95, P =0. 086). The mildest side effect was found in the hypofractionated RT group. No statistically significant differences were found in the patterns of first failure. Conclusion The hypofractionated RT scheme showed potential survival benefits for patients with LS-SCLC and should be considered in the setting of randomized clinical trials.

Objective To investigate whether the change of beam set-up methods will influence the dosimetric quality of intensity modulated radiation therapy (IMRT) for non-small cell lung cancer (NSCLC).Methods Twenty-one stage Ⅰ-Ⅲ NSCLC patients were selected for this study.The technique of step and shoot was used and three different beam set-up methods were chosen for IMRT planning,including IMRT-7 with nine equal-spaced beams angled 0°,51°,102°,153°,204°,255°and 306°; IMRT-5 with five equal-spaced beams angled 0°,72°,144°,216°and 288°; and IMRT-5m which was created from IMRT-7 but excluded 2 fields (51°and 102° were omitted if there was lesion in the right lung,while 255°and 306° were excluded if there was lesion in the left lung).The dose constrains ofnormal lungs for IMRT were set according to V5-V60 of normal lungs obtained from the same patient's actually treated 3D-CRT dose volume histogram.The prescription dose for IMRT started from 65 Gy,and then escalated or decreased step by step by 2 Gy once a time until the best plan was obtained.Results For normal lung dose,IMRT-5m had lower V5-V25 than the other two groups; but there was no significant difference in V30-V40.IMRT-5 was the worst for V45-V60; and mean lung dose was lowest in IMRT-5m.Dose parameters of esophagus and spinal cord,target conformity index,and total monitor units were all similar among difference plans.IMRT-5m had lowest heart V40 compared to the other two groups.For target heterogeneity index,IMRT-5 was higher than IMRT-7,but there were no significant differences among IMRT-5m,IMRT-5 and IMRT-7.Compared to 3D-CRT,the prescription dose could be increased by (5.1 ±4.6) Gy for IMRT-7,(3.1 ±5.3) Gy for IMRT-5,and (5.5 ±4.8)Gy for IMRT-5m.Conclusion Fewer beams and modified beam angles could result in similar,even better plan quality.

Objective To assess the rotational set-up errors in patients with thoracic neoplasms. Methods 224 kilovohage cone-beam computed tomography (KVCBCT) scans from 20 thoracic tumor pa-tients were evaluated retrospectively. All these patients were involved in the research of " Evaluation of the residual set-up error for online kilovohage cone-beam CT guided thoracic tumor radiation". Rotational set-up errors, including pitch, roll and yaw, were calculated by 'aligning the KVCBCT with the planning CT, using the semi-automatic alignment method. Results The average rotational set-up errors were -0.28°±1.52°, 0.21°± 0.91° and 0.27°± 0. 78° in the left-fight, superior-inferior and anterior-posterior axis, respective-ly. The maximal rotational errors of pitch, roll and yaw were 3.5°, 2.7° and 2.2°, respectively. After cor-rection for translational set-up errors, no statistically significant changes in rotational error were observed. Conclusions The rotational set-up errors in patients with thoracic neoplasms were all small in magnitude. Rotational errors may not change after the correction for translational set-up errors alone, which should be e-valuated in a larger sample future.

SCLC can spare more volume of the normal lungs and e-sophagus, and has the ability of dose escalation.