AIM: To determine the inhibition of VEGF small interfering RNA (siRNA) on expression of VEGF protein and SMMC 7721 cell growth. METHODS: Nine VEGFsiRNA sequences with nineteen nucleotides were designed under the assistance of computer. The inhibitory effect of VEGFsiRNA was analyzed by CCK8. The protein level of VEGF in the media was determined by ELISA. The change of cell cycle was detected by flow cytometry. RESULTS: All the VEGFsiRNA were capable of inhibiting the proliferation of SMMC 7721 cells significantly compared with control and lipofectamine control, while the inhibitory effects of VEGFsiRNA2, VEGFsiRNA4, VEGFsiRNA6, VEGFsiRNA7, VEGFsiRNA8 and VEGFsiRNA9 were better than that of antisense oligodeoxynucleotide. All the VEGFsiRNA reduced the expression of VEGF protein. The effect of VEGFsiRNA7 and VEGFsiRNA2 were the best with the inhibitory rates of 52.65% and 50.43%, respectively. VEGFsiRNAs induced the S arrest in SMMC 7721 cells. The signs of apoptosis in SMMC 7721 cells induced by VEGFsiRNA were not observed. CONCLUSION: VEGFsiRNA sequences were designed and synthesized successfully. VEGFsiRNA effectively inhibited the proliferation of SMMC 7721 cells and reduced the level of VEGF protein.

Objective Nine VEGF siRNAs were designed and their efficiencys were evaluated for introducing new and effective design rules of siRNA.Methods Nine VEGF siRNA sequences with nineteen nucleic acids were designed.After construction in vitro,VEGF siRNAs were transferred into neuroglioma cells by lipofectamine.The level of VEGF protein in the media was determined by ELISA 48 hours after transferring.The relationship between the characteristics of VEGF siRNA and its' interfering effect were analyzed.Results A quantitative parameter called H-b index were used to characterize the secondary structure of VEGF mRNA.A low value of the H-b index would mean that the structure of mRNA is loose,in which the nucleotides of the targeting region are likely to be accessible by siRNA and the effects are better.The experiments showed a negative correlation between the suppression of VEGF protein expression and H-b index(r=-0.498).The H-b indexs of siRNAs targeting VEGF mRNA 386～406 and 401～421 position were less,however the effects of suppression were better.The siRNAs conferred a strong RNAi activity with A residue at position 6 and A/U residues at position 19 of sense strand of siRNAs.Thethermodynamic properties of siRNA play a critical role in the transferring effects.The lower average of internal stability values for positions 9～14 of the antisense strand is facilitating the interfering effect.The lower binding energy is the more stable incorporation between antisense strand of siRNA and mRNA present,the more favorable is for RISC cleaving mRNA.Conclusions The sequences of VEGF siRNA have been designed successfully and effectively inhibited the expression of VEGF protein.

Drugs, Chinese Herbal ; therapeutic use ; Glomerulosclerosis, Focal Segmental ; drug therapy ; Humans ; Sclerosis

There is a broad and urgent need for the clinical application of anticancer nanomedicine in tumor therapy, but the complex biological barrier in solid tumors has always been the main obstacle to infiltrating nanomedicine into the tumor. The traditional design of nanomedicine based on enhanced permeability and retention (EPR) effect still has some limitations in tumor permeability, it is urgent to find other design theories. Therefore, this review summarizes two novel strategies, active transcytosis and immune cell-mediated tumor penetration, for promoting tumor penetration of anticancer nanomedicine.

AIM: To construct eukaryotic expression vector of human vascular endothelial growth factor (VEGF) gene, and to explore the effects of VEGF gene on proliferation and chemosensitivity of leukemia cells. METHODS: The recombinant eukaryotic express plasmid pEGFP-C1-hVEGF_(165) was constructed with conventional gene engineering methods. The pEGFP-C1-hVEGF_165 was transfected into leukemia cell K562. The proliferation of the recombinant eukaryotic expression plasmid was analyzed by CCK8 method. The level of VEGF mRNA was detected by reverse transcription and quantitative polymerase chain reaction (RT-PCR). The expression level of VEGF protein was determined by ELISA in cell culture medium and cells. The cell cycle was detected by flow cytometry. RESULTS: VEGF_(165) eukaryotic expression vector was successfully constructed, confirmed by enzyme digestion and sequencing. Compared with the K562 cells transfected pEGFP-C1, the K562 cells transfected with pEGFP-C1-hVEGF_(165) grew faster and expressions of VEGF mRNA and protein increased significantly. In addition, K562 cells transfected with pEGFP-C1-hVEGF_(165) had relatively higher cell survival rate to chemotherapy drugs homoharringtonine(HHT) and fluorouracil(FU). CONCLUSION: VEGF_(165) eukaryotic expression vector increases the level of VEGF mRNA and protein expression, accordingly promotes the proliferation of leukemia cells and decreases the sensitivity of leukemia cells to HHT and FU.

Objective To study the prostate volume of normal children and adolescents and to evaluate the development of prostate central zone and peripheral zone.To provide the preliminary MRI standards of prostate growth and development for nomal Chinese children and adolescents.Methods Five hundred and fifty eight healthy male volunteers,aged from 2 to 25 years were retrospectively analysed.They were divided into 5 groups by the ages (group A:2 to 5 years,group B:6 to 10 years,group C:11 to 15 years,group D:16 to 20 years and group E:21 to 25 years).All the volunteers underwent conventional MRI examinations at our institution (T2WI axial and coronal image acquisition).The prostate volume of different age groups were measured,the development of the central zone and peripheral zone were scored by two radiologists,and the datas were treated by Nonparametric test.Results Group A children's prostate nearly not development,only individual MRI measurable its size; Group B most not development,only 6 children visible prostate form,but can't distinguish the anatomical division; Group C prostate gland development better,MRI can measure the size,but distinguish its central and peripheral area is difficult ; Group D prostate gland development rapidly,differences are obvious,but only a few central and peripheral area boundary clear; Group E the central and peripheral area boundary clear,form full,T2 WI signal contrast clear.The median prostate volumes of 5 groups were 0.000,0.000,2.450,7.990 and 10.600 cm3,respectively(x2 =215.452,P ＜ 0.01).The median prostate scores of 5 groups were 0.04,0.31,1.34,2.23 and 2.60,the prostate volumes were significant difference among these 5 groups except group A and group B(x2 =127.460,P ＜0.01),there were significant difference among the volumes and the scores of these five groups (P ＜0.01).Conclusions MRI provided the basis of objective and truth for the the measurements of volume and the observations of prostate development in this study,that offers preliminary reference range for normal Chinese children and adolescents prostate development status.

Objective To observe the effects of Lentinan(LNT) combined with Cisplatin (DDP) on the apoptosis induction in mice bearing Lewis lung carcinomas (LLC). Methods Lewis lung carcinoma cells (1?107) were subcutaneously inoculated into BALB/c mice. Mice were randomized into control groups and treatment groups. Observe the morphologic views under light microscope (LM) and electron microscope (EM). Results Compared with the control groups, the treatment groups were morphologic characteristic by relatively smaller in size, cell atypia were not evident, pathalogical karyokinesis were less interstitial blood vessel were fewer, while lymphocyte infiltrate into the interstitial. Apoptotic bodies could be seen under EM. Within all treatment groups, LNT+DDP group showed significant difference. Conclusions Lentinan combined with Cisplatin showed significant antitumor effects, and mechanism maybe inducing the tumor cell apoptosis and stimulate the proliferation of lymphocyte.

Objective To observe the clinical efficacy of abdominal acupuncture plus moxibustion in treating cervical spondylosis of vertebral artery type.Method A hundred patients with cervical spondylosis of vertebral artery type were randomized into a treatment group and a control group, 50 cases in each group. The control group was intervened by abdominal acupuncture plus electroacupuncture; while the treatment group was by moxibustion in addition to the treatment given to the control group. After a treatment course, the onset and remission time of vertigo was observed, and the clinical efficacies were compared between the two groups.Result The total effective rate was 96.0% in the treatment group versus 93.8% in the control group, and the difference was statistically significant (P<0.05). There was a significant difference in comparing the onset and remission time of vertigo between the two groups (P<0.05). In comparing the relapse rates 3, 6, and 12 months after the intervention between the two groups, the difference were statistically significant (P<0.01).Conclusion Abdominal acupuncture plus moxibustion is an effective approach in treating cervical spondylosis of vertebral artery type.

Objective To evaluate the roles of B-type natriuretic peptide (BNP) in predicting the severity of community-acquired pneumonia (CAP) by studying the correlation between them.Methods A total of 202 patients with CAP admitted from December 2011 to December 2012 were enrolled in this study.All these patients were checked with laboratory tests for BNP level,C-reactive protein (CRP),white blood cell count (WBC) as well as other markers needed for obtaining pneumonia severity index (PSI).The differences in BNP levels,CRP levels,and WBC were compared among different degrees of pneumonia severity,and the correlation between BNP levels and PSI was investigated by a linear correlation analysis.The patients enrolled were divided into a high-risk group (defined as Ⅳ-Ⅴ grade of PSI) and a low-risk group (defined as Ⅰ-Ⅲgrade of PSI).Meanwhile,they were also divided into a survivor group and a non-survivor group according to outcomes.BNP levels between the two groups were compared,and a receiver operating characteristic (ROC) curve analysis was performed on the BNP levels versus PSI.Results BNP levels increased with CAP severity (r =0.782,P ＜0.01).The mean level of BNP (263.2 ± 119.6) pg/mLof patients in the high-risk group was significantly higher than that of patients (71.5 ± 54.3) pg/mL in the low-risk group (P ＜ 0.01).The patients in the non-survivor group had significantly higher BNP levels compared to the survivor group [(343.86 ± 125.49) vs.(183.00 ±121.71) pg/mL,P ＜ 0.01].In addition,there were positive correlations between BNP levels and PSI (r =0.782,P＜0.001),between BNP levels and CRP levels (r =0.560,P＜0.01),and between BNP levels and WBC (r =0.513,P＜0.001).The BNP level had a high accuracy in predicting the severity of CAP (AUC =0.952).The optimal cut-off point of BNP level for distinguishing high-risk from low-risk patients was 125.0 pg/mL,with a semitivity of 0.891 and a specificity of 0.946.Moreover,BNP level had a definite accuracy in predicting mortality (AUC =0.823).Its optimal cut-off point for predicting death was 299.0 pg/mL,with a sensitivity of 0.675 and a specificity of 0.816; its negative predictive cut-off value was 0.926,and positive predictive value was 0.426.Conclusions BNP level is positively correlated with the severity of CAP.The patient with BNP level above 125.0 pg/mL should be hospitalized immediately and patients with BNP level higher than 299.0 pg/mL are at the high risk of death.Therefore,BNP is a useful biomarker for evaluating the severity of patients with CAP.

Drug Interactions ; Humans ; Proton Pump Inhibitors ; pharmacology ; Ticlopidine ; analogs & derivatives ; pharmacology