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 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.

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 investigate the effects of Guiqi polysaccharide (GQP) on the telomerase activity in premature senescence of WI-38 cells;To discuss its mechanism of action.Methods MTT assay was used to observe the effects of different concentrations of H2O2 on the proliferation of WI-38 cells and screened the best concentration of H2O2 to induce premature senescence cellular model in WI-38 cells. ELISA and chemical staining method were used to evaluate the protection of GQP in telomerase activity and senescence-associated β-galactosidase activity in premature cellular senescence.Results 200μmol/L H2O2 treatment could induce premature senescence in WI-38 cells. There were no significant differences in senescence-associated β-galactosidase and telomerase activity between GQP group and normal control group (P>0.05), but had significant difference with model group (P<0.01).Conclusion GQP can increase the telomerase activity and inhibit the senescence-associated β-galactosidase activity in premature senescence of WI-38 cells.

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 .

Objective Cerebral microbleeds (CMBs) are important indicators of cerebral small vessel disease .However, it is still unclear whether endothelial dysfunction is involved in CMBs .The aim of this study is to investigate the correlation between CMBs and soluble E-selectin (sE-selectin) in patients with acute ischemic stroke . Methods Based on the results of MRI (3.0 T) susceptibility weighted imaging , we divided patients with first acute ischemic stroke into a CMBs group ( n=63 ) and a non-CMBs group (n=63), and recruited another 45 volunteers with normal MRI findings as controls .We collected and conducted comparative a-nalysis on the demographic data , biochemical variables ( including the sE-selectin level ) , vascular risk factors , and the number of CMBs of the patients . Results Ordinal logistic regression analysis showed a significantly positive correlation between sE -selectin and the number of CMBs (OR=1.062, 95%CI:1.023-1.103, P=0.002), higher systolic blood pressure associated with more CMBs (OR=1.014, 95%CI:1.002-1.025, P=0.021). Conclusion Serum sE-selectin is significantly positively correlated with and can be used as a biological marker for the severity of CMBs .

Background and purpose:We investigated whether lfuorine-18 lfuorodeoxyglucose (18F-FDG) maximal standard uptake value (SUVmax) of the primary tumor (SUV-T), SUVmax of the regional lymph nodes (SUV-N) or the overall loco-regional lesion SUVmax (SUV-TOTAL) was related to survival of patients with stage Ⅲ non-small cell lung cancer (NSCLC) who received Cetuximab and combined definitive chemoradiotherpay. Methods:From September 2009 to July 2012, seventeen patients with unresectable stageⅢNSCLC receiving cetuximab with cisplatin/vinorelbine (NP) followed by concomitant NP and intensity-modulated radiotherapy (IMRT) at the Fudan University Shanghai Cancer Center were enrolled onto a prospectively study. All patients received positron emission tomography/computerized tomography (PET/CT) scans within 2 weeks before enrolment. Univariate analysis were used to assess the correlation between SUV-T, SUV-N, SUV-TOTAL, gender, age, histology, tumour-node-metastasis (TNM) stage, performance status (PS) as well as smoking status and survival. The factors which showed statistical signiifcance entered into multivariate Cox-regression model. Survival functions of different populations were estimated by Kaplan-Meier method and compared by Log-rank test. Results:In the univariate analysis, SUV-T, SUV-N, SUV-TOTAL, PS and smoking status were prognostic factors. The best cut-off values for SUV-T, SUV-N and SUV-TOTAL were 11, 11 and 20, respectively. Multivariate analysis revealed that SUV-TOTAL (P=0.012), SUV-T (P=0.025), and SUV-N (P=0.033) were independent predictors of survival with hazard ratio (HR) of 14.7, 11.2, and 6.2, respectively. Conclusion:Local, regional and locoregional maximal SUVs deifned by 18F-FDG PET-CT scanning may have a strong correlation with survival in this patients setting, which merits further study.

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.

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.