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 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 .

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.

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 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 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 explore the localization of epileptogenic focus and select the appropriate surgical procedures for post-traumatic epilepsy. Methods The clinical data of 21 patients with post-traumatic epilepsy were studied retrospectively. Epileptogenic focus was located by comprehensively analyzing data of electro-neurophysiology, neurological imaging and clinical manifestation. Surgical procedures were performed in all patients, including resection of lesion and peripheral cortex in 12 patients, epileptogenie focus resection plus low power bipolar coagulation in five, anterior temporal iobectomy plus amygdalohippocampectomy in three and corpus callosotomy in one. Results All patients were followed up from 6 months to 3 years, which showed satisfactory outcome in eight patients, marked improvement in six, improvement in five and slight improvement in two. The total effective rate was 90%. Conclusions Surgical procedure is important for intractable post-traumatic epilepsy. The good efficacy depends on precise localization of epileptogenic focus and combined application of various surgical procedures.

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.