Objective To replace the endogenous CTL epitope LLD in HBsAg with EYILSLEEL of glypican(GPC3)in order to prepare a GPC3-HBsAg multiple peptides vaccine for HBV infection.Methods The HBsAg/GPC3 DNA was amplified by gene splicing by overlap extension(SOEing)PCR from pcHBsAg plasmid and then inserted into pBSSK+ vector to construct a pBSSK/GPC3 vector.The vector was then identified by PCR,double digesting and sequencing.The fragment encoding GPC3 CTL epitope EYILSLEEL was obtained by double digesting the plasmid pBSSK/GPC3 and then inserted into pcDNA3.1+ vector.Results Sequencing and restrict endonulease digestion indicated that eukaryotic expression vector pcDNA-HBsAg/GPC3 was constructed successfully.Conclusion The endogenous CTL epitope LLD of HBsAg is replaced by the EYILSLEEL,and an eukaryotic expression vector pcDNA-HBsAg/GPC3 is constructed.

BACKGROUND:A large number of experiments have shown that bone marrow mesenchymal stem cels (BMSCs) have good effects on the treatment of lung disease or improvement of lung injury, and its therapeutic effect is mainly oriented to reduce the inflammatory response. OBJECTIVE: To study the effects of BMSCs combined with reduced glutathione in murine models of bleomycin-induced lung injury. METHODS:BMSCs were isolated from a male NOD/SCID mouse and cel morphology and phenotype were observed. Sixty-four female NOD/SCID mice were randomly divided into a control group, a model group, a BMSCs group and a BMSCs+reduced glutathione group (combined group). The control group received intratracheal injection of normal saline, the model group received intratracheal injection of bleomycin, and the BMSC group received BMSCs injectionvia the tail vein at 2 hours after intratracheal injection of bleomycin, and combined group received BMSCs+reduced glutathione injection via the tail vein at 2 hours after intratracheal injection of bleomycin. Al mice were kiled after 7 days, and levels of tumor necrosis factor-α, interleukin-β, malondialdehyde in lung tissue were detected and lung tissue specimens were obtained for pathological examination. RESULTS AND CONCLUSION:BMSCs were fibroblast-like cels, Positive for CD10, CD13 and CD44, but negative for CD34 and CD45. Compared with the control group, the levels of tumor necrosis factor-α, interleukin-β and malondialdehyde were increased in the model group, while decreased significantly in the BMSCs group and combined group (P < 0.05), especialy in the combined group (P < 0.05). Pathological examination showed that more serious lung injury was observed in the model group and BMSCs group than the combined group. These findings indicate that BMSCs combined with reduced glutathione can more effectively protect against bleomycin-induced lung injury.

Objective To evaluate the therapeutic effects and adverse events on treatment of primary premature ejaculation (PE) with paroxetine hydrochloride and/or tamsalosin .Methods 225 cases of healthy men ,a history of lifelong PE and an intravaginal e‐jaculation latency time(IELT) < 120 sec were included in this study .The patients were divided into three groups(with 75 cases in each group) .Group A were given paroxetine hydrochloride 20 mg/d for 8 weeks ;group B were given tamsalosin 0 .2 mg/d for 8 weeks ;group C were given paroxetine hydrochloride and tamsalosin for 8 weeks .The effects and adverse events were evaluated by the overall change and fold increase in average IELT and the mean change in all four measures of the premature ejaculation profile (PEP) .Results The reliable data from 198 patients were achieved .The mean IELT after treatment were significantly improved in all groups than that of before treatment (P<0 .05) .The mean IELT in group C was increased for 8 .15 min after treatment .The in‐creased folds of mean IELT in group C (8 .02 folds) was significantly higher than that in group B (1 .98 folds)and group A (6 .92 folds)(P<0 .01) .The mean PEP scores that include measures of perceived control over ejaculation ,satisfaction with sexual inter‐course ,ejaculation‐related personal distress ,ejaculation‐related interpersonal difficulty were significantly improved in all groups after treatment .The mean PEP scores in group C had more significant improvements than that in group A and B .The prevalence of ad‐verse events in group A ,B ,C were 10 cases(13 .3% ) ,2 cases(2 .6% ) and 9 cases(12 .0% ) ,respectively .Conclusion Paroxetine hydrochloride combined with tamsalosin lead to better therapeutic effects and could be a priority selection for the treatment of pri‐mary PE .

Objective To explore the value of two dimensional and color Doppler flow imaging(2D CDFI)for the diagnosis and differentiation of the carotid body tumor.Methods Retrospective analysis was made on 12 patients with carotid body tumor examined by 2D CDFI,and differentiations were conducted between several similar diseases.All the cases were confirmed by surgical pathology.Results The carotid body tumor in 2D CDFI was shown as follows: Low echoic substantial masses were found in the carotid branch with clearly boundary,and or partly outline.Most in small size were located at the branch of common carotid artery,so the interval between external and internal carotid arteries was enlarged.The masses in big size always grew around common cartid artery internal carotid artery and external carotid artery.More color flow signals were detected in the masses,and most were artery flow.The relationship between the mass and the carotid was shown clearly by 2D CDFI.Conclusions 2D CDFI is a valuable and practical method in diagnosis of carotid body tumors.

BACKGROUND: Comparing with diabetic nephropathy and diabetic retinopathy, diabetic cardiomyopathy is put forward exceeded later and the research about it was not enough. It is difficult to detect diabetic cardiomyopathy in the earlier period so that it has usually been neglected.OBJECTIVE: To evaluate features of dysfunction of left ventricle in early diabetic cardiomyopathy by Doppler tissue imaging.DESIGN: Completely randomized and controlled experiment.SETTING: Ultrasound Department of General Hospital of Chengdu Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out at Ultrasound Department of General Hospital of Chengdu Military Area Command of Chinese PLA from March to June 2002. Totally 56 New Zealand rabbits of either gender were selected.METHODS: ① Ten New Zealand rabbits was pulled out randomly from 56 as control group. The rest of 46 New Zealand rabbits were replicated diabetes mellitus by Streptozotocin (STZ), rabbits were absolute diet 18 hours and injected intravenously to ear marginal vein with 65 mg/kg of STZ. Then these rabbits were randomly divided into 4 groups as 2, 4, 6, 8th experimental group, with 10 rabbits in each group. ② Digit ultrasonic cardiogram equipment was used to examined interventricular motion of mitral annulus at 2nd,4th, 6th and 8th week in model groups and comparison group. The parameters such as systolic peak velocity (Sa), preejection period (PEP), local ejection time (ET), systolic mean velocity (Vm) of four various portions in mitral annulus and diastole early period velocity (Ea), diastole later period velocity (Aa), Ea/Aa were determined. ③ SPSS 11.0 was used for statistical analysis,one-factor analysis was adopted to compare difference of groups.MAIN OUTCOME MEASURES: The motion of lateral wall and posterior interventricular septum at mitral annulus were estimated by PW-DTI.RESULTS: Among 56 rabbits, 50 entered the final analysis and other 6 were lost because of failure modeling. Diastole dysfunction of left ventricle was discovered in STZ-inducod diabetic rabbits detected by tissue Doppler imaging at the 4th week. It showed that not only the tite of early period of diastole peak velocity (Ea) in mitral ring lateral wall and post-interval of left ventricle was reduced, but also the tite of Ea/Aa was reduced notably (P ＜ 0.05). Systolic dysfunction of left ventricle was discovered in STZ-in-duced diabetic rabbits detected by tissue Doppler imaging at the 6th week.It showed that local contraction peak velocity (Sa) of lateral wall and postseptum of mitral ring was decreased strikingly (P ＜ 0.05). Mean systole velocity of four various parts of mitral ring (Vm) was reduced at the 6th week (P ＜ 0.05). Pre-ejection period time (PEP) at post-septum and ejection time (ET) was elongated at lateral wall at the 8th week (P ＜ 0.05).CONCLUSION: ① Diastolic dysfunction of left ventricle is characterized by decrease of early period of diastole peak velocity and the tite of Ea/Aa.② Systolic dysfunction of diabetic cardiomyopathy is characterized by decrease of local contraction peak velocity and elongation of pre-ejection period time and ejection time.

Objective To assess potential suitability of the intercostal artery as a conduit in coronary bypass graft.Methods Collected 36 cases of chinese adult thoracic aorta CTA original cross-sectional images,the images were retrospectively reconstructed under volume rendering technology (VR),multi-planar reconstruction (MPR),curved planar reformation (CPR) and maximum intensity projection(MIP).Observed and measured the data of the 8th,9th and 10th bilateral intercostal arteries and internal thoracic arteries.Results Toshiba 320-slice volume scanning CT can clearly display the intercostal arteries.The average length and diameter at corresponding point of the left and right 8-10th intercostal artery is similar,all the length is longer than 180 mm,and the diameter at axillary line is all bigger than 2.1 mm,the mean length of right intercostal arteries is slightly longer than the left intercostal arteries,the diameter at each corresponding point of left and right intercostal artery is similar,and equivalent to approximately 85％-90％ of internal thoracic artery.Conclusion Intercostal artery can be used as an ideal alternative vessel in coronary artery bypass grafting.

Objective To study the effect of the uncertain deflection of the Delta4 phantom (ScandiDos AB,Sweden) in setting up on the Gamma index passing rate during the VMAT plan verification.Methods Two patients with head and neck cancer,two with lung cancers and one with pelvic cancer receiving VMAT radiotherapy were randomly chosen.By means of Eclipse8.6 TPS the treatment plans elaborated for the five patients were picked up to make the verification plans and Delta4 was used to perform dose verification On VARIAN Clinac Ⅸ.The Delta4 phantom was precisely set up first,and then it was deflected in a given angle towards the horizontal direction in relation to the center of the linear accelerator isocenter to perform the dose verification for 11 times successively.To figure out the relationship between the deflection angle of the Delta4 phantom and the Gamma index passing rate.Results As the Delta4 phantom was deflected by 0.0°,0.2°,0.4°,0.6°,0.8°,1.0°,1.2°,1.4°,1.6°,1.8° and 2.0° in sequence,the measured Gamma index passing rates presented a slight decline,but all greater than 90％ (DD 3％,DTA 3 mm).Conclusions In the VMAT plan verification,the Gamma index passing rate of Delta4 has no dependence on the uncertain deflection of the Delta4 phantom provided that the uncertain deflection of the Delta4 phantom is no greater than 2°,but the passing rates of DD and DTA vary significantly with the uncertain deflection of the Delta4 phantom.

Objective To investigate the molecular mechanisms of synergistic effects of BFA and CDDP on human lung cancer GLC-82 cells, and to test the levels of PERK-ATF4 pathway. Methods GLC-82 cells were incubated with 50 ng/mL of BFA or/and 2 μg/mL of CDDP for 24 or 48 hours. The levels of PERK, p-PERK and ATF4 in GLC-82 were analyzed by real-time PCRand/or Western Blot. Results The levels of PERK were lowest in CDDP group, but higher in BFA group (P < 0.05), the highest in group of BFA+CDDP (P < 0.05 or P < 0.01). The p-PERK level decreased in group of BFA+CDDP (P < 0.05 or P < 0.01). There was no significant change of ATF4 expression in CDDP group, but ATF4 expression increased slightly in BFA group, and increased further in group of BFA+CDDP (P < 0.05 or P < 0.01)which was also higher than that in BFA group or CDDP group (P < 0.05 or P < 0.01). Conclusions The upregulated levels of PERK and ATF4 by the combination of BFA and CDDP may be one of the mechanisms of synergistic anti-cancer effect of BFA and CDDP on GLC-82 cells.

Objective To explore more satisfactory results in correcting the aesthetically short nose combined with low and blunt nasal tip by using autologous cartilage grafts.Methods External rhinoplasty approach was preferred.Nasal septal cartilage and auricular cartilage or rib cartilage of patients were adopted,as necessary,various shapes of grafts were carved,such as septal extention graft,columella strut and shift graft.Then we used these autologous cartilage grafts with suture and removal techniques to correct the aesthetically short nose combined with low and blunt nasal tip.The length of the nose (n-prn) and the projection of nasal tip (sn-prn) were measured before and after the operation.Paired t-test was adopted to evaluate the results.Results Thirty-one patients accepted the nasal tip rhinoplasty with autologous cartilage whose nasal tip was over rotation,blunt and low.All the patients were followed up for 3 months.Thirty patients satisfied with the results,accounting for 96.7 ％ of the total.()ne (3.3 ％) patient dissatisfied with the result because of postoperative asymmetry nostrils.Nose length before and after surgery was significantly different (P＜0.05).Nasal tip projection before and after surgery was also significantly improved (P＜0.05).Conclusions It is an effective method to use autologous cartilage graft for correcting the aesthetically short nose combined with low and blunt na sal tip,with low complications.