Objective:To identify the expression of plasmid pLNCX-EGFP-C1-hri targeting the gene of Human ribonuclease inhibitor (hri) in PA317 cells which is capable of expression in mammalian cells.Methods: The vector of pLNCX-EGFP-C1-hri was transfected into PA317 cells by Lipofectamine 2000 and then the expression of recombinated plasmid was verified in living cells by observing the transcription level of egfp-hri fusion gene mRNA with RT-PCR method and the expression level of egfp-fusion hRI protein with western blotting method respectively.Results: Both RT-PCR and western blotting showed the egfp-hri fusion gene was obviously expression in PA317 cells.Conclusion: The plasmid of pLNCX-EGFP-C1-hri targeting hRI is successfully constructed and the protein of hRI can be expressed in PA317 cells correctly.

OBJECTIVE:To study the compatible stability of Levofloxacin with Tinidazolc in Glucose Injection.METHO-DS:The contents were determined by UV-spectrophotometry and the appearance and pH value were observed within 8h after mixing of levofloxacin with tinidazolc in glucose injection at room temperature(20℃).RESULTS:The contents,pH value and appearance of the mixed solution showed no significant changes within 8h.CONCLUSION:The mixture of the levofloxacin and tinidazolc in glucose injection can be used within 8h after mixing.

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

OBJECTIVE:To prepare dexamethasone acetate borneol cream and to establish its quality control method.METHODS:Dexamethasone acetate borneol cream was prepared by grinding method;dexamethasone acetate and phenol were identified by TLC,and the content of the principal agent was determined by ultraviolet spectrophotometry.RESULTS:The prepared cream fitted the description of China Pharmacopeia(2005 edition) in identification and tests etc.The linear range of dexamethasone acetate was 12.06～28.14 ?g?mL-1(r=0.999 7),and the average recovery rate of 98.52%(RSD=0.66%).CONCLUSION:The preparation is reasonable in compounding,simple and feasible in techniques,and its quality is stable and controllable.

Complement system is an important component of innate immunity and has been recognized as an effective means to inhibit tumor. However,in last decades,accumulating studies showed unexpected results that the complement components and their activation products could promote development of malignancies by secreting growth factors,activating signal pathway and promoting tumor angiogenesis. Herein,the relationship between the complement system and tumor is reviewed.

Objective To investigate the biological effects of internal radiation and therapeutic effectiveness of 131I-labeled anti-epidermal growth factor receptor (EGFR) in colorectal cancer of model mice.Methods Nano-liposome characterized for EGFR-targeting was constructed.The efficacy of cellular binding and uptake of the liposome was evaluated by the analysis of confocal microscopy observation and the iodide uptake assay.After intra-tumor injections of 74 MBq (740 MBq/ml) 131 I-antiEGFR-BSA-PCL,131 I-BSA-PCL,131 I or an equivalent volume of normal saline.The biological effects of internal irradiation and therapeutic efficacy of the liposomes on colorectal cancer modeled in a male BALB/c mouse were evaluated by means of tumor size,body weight,histopathology,and SPECT imaging.Results The confocal fluorescence images showed that the antiEGFR-BSA-PCL was successfully internalized into LS180 cells.The 131I uptake efficacy of 131I-antiEGFR-BSA-PCL was significantly higher than that of 131I-BSA-PCL in LS180 cells (t =2.77-5.40,P ＜ 0.01).Tumor size measurement showed that tumor growth was inhibited by the treatment with 131 I-EGFR-BSA-PCL and 131I-BSA-PCL,but had no significant differences between these two groups (P ＞0.05).It was found that the 131I-antiEGFR-BSA-PCL was markedly taken up by the tumor and reac hed its uptake value of (21.61 ± 1.0 1) and (20.58 ± 0.65)％ ID/g at 72 h following drug injection,which was higher than the uptake value of 131 I (t =9.36,8.69,P ＜ 0.01).SPECT imaging assay showed that,after being injected into mouse tumor,the 131 I-EGFR-BSA-PCL and 131I-BSA-PCL were uniformly distributed inside the tumor.Conclusions 131 I-antiEGFR-BSA-PCL obviously suppresses the development of colorectal cancer in mice.

Objective To construct 131 I labeled anti?EGFR immunoliposome nanoparticle ( 131 I?Cetuaximab ( C225)?BSA?PCL) , and investigate its inhibitory effect on EGFR?overexpressing cancer cells in vitro. Methods Anti?EGFR liposome nanoparticle C225?BSA?PCL and non?targeted liposomes BSA?PCL were constructed. The products were observed with transmission electron microscopy and dynamic light scat?tering. The EGFR?targeted binding and cellular uptake in EGFR?overexpressing cancer cells were observed with flow cytometry and confocal microscopy. Anti?EGFR and non?targeted liposomes were labeled with 131 I using the chloramine?T method. The targeted cell killing effects of 131 I labeled liposomes were analyzed using MTT assay. The time?dependent cellular uptake analysis was used to evaluate the slow?release effects of the 131 I labeled liposomes. The independent?samples t test was used for data analysis. Results The EG?FR?targeted liposome C225?BSA?PCL and non?targeted liposome BSA?PCL were successfully constructed, and the effective diameters were approximately 130-180 nm. Flow cytometry and confocal microscopy re?vealed significant uptake of C225?BSA?PCL in EGFR?overexpressing tumor cells. BSA?PCL could also bind to cells with minimal and weak tumor retention. The EGFR?targeted radioactive liposome 131I?C225?BSA?PCL showed greater targeted cell killing effect than non?targeted liposome 131I?BSA?PCL,the IC50 values of 131I?C225?BSA?PCL and 131 I?BSA?PCL were 0. 03-1. 32 and 0. 25-12. 19, respectively. The uptakes of 131 I?C225?BSA?PCL was higher than that of 131 I?BSA?PCL ( t=3.03-16.86, all P<0.05) and reached the maxi?mal level at 4 h after incubation. Conclusions The EGFR?targeted liposome C225?BSA?PCL demonstrated superior cellular binding and uptake on EGFR?overexpressing cancer cells compared with BSA?PCL. The EGFR?targeted radioactive liposome 131 I?C225?BSA?PCL had favorable intracellular retention and excellent targeted cell killing effect, and could effectively suppress the growth of EGFR?overexpressing cancer cells.

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.

Background Light stimulation at different wavelength influences the development of eyes.It has been showed that blue light can inhibit the growth of eyeball.To study whether blue light exposure can delay the extension of myopia is an interested research project.Objective This study was to investigate the effect of blue light with short wavelength on ocular growth in form deprived myopia (FDM) in guinea pigs and provide a new option for the prevention and treatment of myopia.Methods Thirty-six 2-week-old guinea pigs were reared in the environment of white light.The right eyes of the animals were occluded to establish the FDM models.The models were randomized into the deocclusion + blue light exposure group,simple deocclusion group and continuous occlusion group according to the random number table.The right eyes of the models were deoccluded for 1 hour per day to give the blue light (430 nm) irradiation in the deocclusion + blue light exposure group,and the right eyes were deoccluded for 1 hour per day only in the simple deocclusion group.In the continuous occlusion group,the right eyes of the models were occluded until the end of this experiment.Anterior chamber depth (ACD),lens thickness (LT) and vitreous cavity depth (VCD) were measured by A-type sonography.The binocular diopter of the guinea pigs was detected using retinoscopy in the mydriatic condition.In the fourth week after experiment,the retinal sections were prepared for the regular histopathological examination,and the scleral tissues next to 1 mm from optical nerve were exacted to obtain the dry weight of scleral tissues.Results In the right eyes of the animals,no significant differences were found in the diopter,ACD,LT and VCD before experiment among the 3 groups (all at P＞0.05).At the end of experiment,the refraction of right eye in the deocclusion + blue light exposure group,simple deocclusion group and continuous occlusion group was (+1.11±0.17)D,(+0.90±0.15)D and (-2.73±0.19)D respectively,with a significant difference among them (F=1 445.470,P=0.000).The VCD in the three groups was (3.70±0.09) mm,(3.78±0.11) mm and (3.91 ± 0.08) mm,respectively,showing a significant difference (F =13.243,P＜0.01).In addition,the dry weight of sclera tissues was (0.61 ±0.09)mg in the deocclusion + blue light exposure group,(0.54± 0.08)mg in the simple deocclusion group and (0.43 ± 0.07)mg in the continuous occlusion group,with a significant difference among the 3 groups (F=10.458,P＜0.01).However,there were no significant differences in the ACD and LT among the 3 groups (F=0.203,0.084,both at P＞0.05).Moreover,in the left eyes,no significant differences were found in the diopter,ACD,LT and VCD before experiment among the 3 groups (all at P＞0.05);while at the end of the experiment,the diopter of the continuous occlusion group was significantly lower than that of the deocclusion + blue light exposure group and simple deocclusion group (all at P＜0.05).No significant differences were seen in the ACD,LT,VCD and dry weight of sclera among the 3 groups (all at P＞0.05).Retinal structure was normal in the left eyes of various groups.However,the retinas were thinner in the right eyes of the deocclusion + blue light exposure group with clear layers; while atrophy of the outer segment of photoreceptor and disorder of cell arrangement were seen in the right eyes of the continuous occlusion group.Conclusions During sensitive period of visual development,blue light stimulation can arrest the extension of posterior sclera and elongation of vitreous cavity,which restrains development of myopia.This blue light at the wavelength of 430 nm is safe to retina.

Objective To evaluated the effect of hyperbaric oxygenation therapy(HBO)on the RhoA ex- pression and nerve function after transient focal cerebral ischemia in a rat model of middle cerebral artery occlusion. Methods One hundred and twenty-six healthy Sprague-Dawley rats were used and randomly divided into a sham op- eration group(shame group,n=42),a treatment group(n=42),and a control group(n=42).The animal model of middle cerebral artery occlusion(MCAO)was established by using the Zea-Longa method with the animals in the treatment and the control groups,and sham operation was performed with those in the sham group.HBO was applied to the animals in the treatment group.The RhoA protein expression was observed by using immunohistochemistry technique,and the neurological function was evaluated by Bederson's scale at different time points after MCAO.Re- sults(1)Weakly positive expression of RhoA could be located in bilateral cortex and the basal ganglia in the sham group.The expression of RhoA in the treatment group and control group was increased as early as 6 hours after MCAO when compared with that of the sham group,and peaked at 48 h after MCAO and decreased after then,but was still higher than that of the shame group at 7th day to 14th day after MCAO.It was also found that the expression of RhoA of the treatment group was significantly lower than that of the control group(P