Objective To investigate the feasibility of recombinant adeno-associated virus encoding secrected forms of human acidic fibroblast growth factor transfecting endothelial progenitor cells.Methods sp-haFGF was obtained through combining signal peptide sequence of FGF-4 with native aFGF gene by PCR.sp-haFGF was cloned into AAV vector plasmid pAAV-IRES-hrGFP.Recombinant AAV encoding sp-haFGF was packaged through co-transfecting HEK293 cells with plasmid sp-haFGF-pAAV-IRES-hrGFP,pAAV-RC and pHelper.Ex vivo cultured EPCs were infected with concentrated rAAV.Expression of green fluorescent protein(GFP) in infected EPC was observed by fluorescence microscope and expression of sp-haFGF in EPCs was verified by RT-PCR and western blot analysis.Results Recombinant AAV encoding sp-haFGF was obtained.After EPCs being infected with rAAV,green fluorescence was found in about 20～30% EPCs,gene of(560 bp) was generated from EPCs by RT-PCR method,and(sp-haFGF) protein was detected in infected cells.Conclusion EPC was efficiently infected by rAAV encoding(sp-haFGF) and(haFGF) was expressed by EPCs,which establishes a basis for therapeutic angiogenesis by transgenic EPCs transplantation.

AIM: To isolate targeted the peptides that binding and internalizing into endometrial carcinoma cell line EAC. METHODS: The tumor cells were screened five rounds of whole cell screen through the Ph.D.-12 phage display library. Analysis of the displayed peptides specific binding efficiency to the tumor cells was proceeded. The DNA of phages was extracted, sequenced and translated to the sequences of amino acid and analysis using computer software. RESULTS: After five biopannings, the isolated phages showed high specificity and strong affinity for their cognate cell types relative to different cell lines. Through sequencing, the sequences of displayed peptides were obtained. CONCLUSIONS: Whole cell screening against EAC cells through random phage peptide library can obtain phage peptides with a highly tissue specific binding and internalizing ability. The peptides do provide a basis for tumor's targeted delivery as therapy vector.

Objective To observe the effects of modified Jianpi Yishen Decoction on urinary osteopontin (OPN) in calcium oxalate nephrolithiasis patients after percutaneous nephrolithotomy (PCNL) or ureteroscope lithotomy (URL);To clarify the mechanism of modified Jianpi Yishen Decoction on the prevention of calcium oxalate kidney stones. Methods Totally 116 calcium oxalate nephrolithiasis patients were randomly divided into trial group (62 cases) and control group (54 cases). The trial group took modified Jianpi Yishen Decoction every other day, while the control group took potassium sodium hydrogen citrate granules three times a day. The concentrations of OPN, urinary calcium and urinary oxalic acid of the patients in the two groups were observed before treatment and 2 weeks and 4 weeks of treatment. Results The concentration of urinary OPN of 2 weeks and 4 weeks of the treatment in the trial group was significantly increased compared with before treatment (P<0.05), and the difference was statistically significant compared with the control group (P<0.05). The concentration of urinary OPN in the control group had no significant change after treatment (P>0.05). The differences in the concentrations of urinary calcium and urinary oxalic acid of the two groups between before and after treatment were not significant (P>0.05). Conclusion Modified Jianpi Yishen Decoction can effectively restrain the formation of the calcium oxalate stones by increasing the level of urinary OPN, which demonstrates effective prevention in the calcium oxalate nephrolithiasis patients after PCNL or URL.

AIM: To study the role of androgen receptor (AR) in hormone-dependent and hormone-independent prostate cancer cell proliferation by knocking down AR expression with adenovirus-delivered siRNA. METHODS: Four well-designed siRNAs were synthesized and inserted into the adenovirus plasmid pShuttle-H1-Ri. The recombinant pShuttle-H1-Ri-AR plasmid was then co-transfected with pcDNA-AR to HEK293 cell line and Western blot was used to detect the inhibitory efficiency of different siRNAs on AR expression. Recombinant adenovirus containing more efficient siRNAs were prepared and used to infect three different humane prostate cancer cell lines including LNCapC4-2B and CWR22Rv1. The efficiency of knocking down AR expression was detected by Western blot. The effect of AR-knocking down on cell proliferation was detected by MTT colorimetric assay. RESULTS: All of the four designed siRNAs could knock down AR expression in transient co-transfection. Infecting with recombinant adenovirus containing more efficient siRNAs in hormone-dependent and hormone-independent prostate cancer cell lines specifically knocked down AR expression with high efficiency. Knocking down AR expression significantly decreased the proliferation rate in all these prostate cancer cells. CONCLUSION: The suppressed expression of AR in prostate cell lines mediated by siRNA could efficiently inhibit the cell proliferation, and these results show that AR plays an important role in the proliferation of hormone-dependent and hormone-independent prostate cancer cells. AR is an important therapeutic target for the treatment of prostate cancer.

OBJECTIVE Toobservetheeffectoftemozolomide(TMZ)incombinationwithtetran-drine(TET)on cell viability,colony formation,migration and cell apoptosis of human glioblastoma U87 cells.METHODS TheviabilityofU87cellstreatedwithTET(8-64μmol·L-1),TMZ(50-400 μmol·L-1 )and TMZ combined with TET (3.2,6.4 μmol·L-1 )was detected by cytotoxicity assays with Cell Counting Kit-8 (CCK-8),the colony formation was detected by Giemsa staining,cell migration ability was detected by Transwell migration assay,cell apoptosis was assayed by flow cytometry using Annexin Ⅴ /PI double staining,and the expression of apoptosis-related proteins expression was detec-tedbyWesternblotting.RESULTS ThedataofCCK-8showedthatTET(r=0.903,P<0.05)orTMZ (r=0.995,P<0.05)could inhibit U87 cell viability alone in a concentration-dependent manner.The cell viability inhibition rate of U87 cells by TMZ co mbined with TET was higher than by TMZ or TET alone. Data showed that the effect of TMZ combined with TET was additive.TMZ 100 μmol·L-1 inhibited U87 cell colony formation and migration ablility compared with normal control.The inhibition rate of U87 cells by TMZ 100 μmol·L-1 combined with TET (3.2 and 6.4 μmol·L-1 )was more significant than by TMZ alone (P<0.05).Compared with TMZ alone,TMZ combined with TET (3.2 and 6.4 μmol·L-1 )signifi-cantly down-regulated the expression of anti-apoptotic protein Bcl-XL,but significantly up-regulated the expression of cleaved caspase 3 protein and cleaved poly(ADP-ribose)polymerase.CONCLUSION TET combined with TMZ can inhibit U87 cell viability,colony formation and migration by activating caspase-dependent apoptotic pathway,resulting in apoptosis.

AIM and METHODS:The Ph.D.-7 phage display library was used to isolate peptides specific for glioma SWO-38 cell by whole cell screening.Moreover,binding efficiency analysis was carried out to test the binding specificity of the clones obtained. RESULTS: After three rounds of biopanning,a high concentration of phage clones was obtained and two of them were found to be highly specific to glioma SWO-38. CONCLUSION: Highly specific clones against neurtral glioma cells can be obtained from a phage display library by simple procedures.

Objective To study of zoledronic acid in the treatment of multiple myeloma bone dis-ease clinical effect and detection of serum macrophage inflammatory protein (MIP)changes of primary mye-loma (mm)in patients with serum macrophage inflammatory protein levels and multiple myeloma bone dis-ease curative effect.Methods 48 cases of multiple myeloma bone disease patients were treated with VTD regimen chemotherapy were randomly and equally divided into two groups,one group (group A)chemother-apy intermission applied zoledronic acid 4 mg per month 1 time,treatment 2 course of treatment,observa-tion of curative effect and adverse reaction,another group (B group)declined to azole phosphonic acid treatment.Results Group of pain Solution of 16 cases were markedly effective,effective in 4 cases,4 ca-ses were ineffective,efficiency 83.3%.B group bone pain relieved markedly effective in 12 cases,effective in 4 cases,8 cases were ineffective,have efficiency 66.7%.A compared to the B,the curative effect was obvious (P <0.05).By enzyme linked immunosorbent assay for the detection of the patients with a,levels of peripheral serum MIP-1a and MIP-1 beta B two groups before and after treatment.Conclusions zole-dronic acid in the treatment of multiple myeloma bone disease effectively,can significantly improve the qual-ity of life in patients with MM patients serum MIP-1a and MIP-1 beta level and multiple myeloma tumor bone disease curative effect is negative correlation,used for evaluating the effect The reference index.

Objective:To investigate the correlation of fluid-attenuation inversion recovery (FLAIR) vascular hyperintensity (FVH) and clinical outcome in patients with middle cerebral artery M1 occlusive stroke.Methods:Patients with acute middle cerebral artery M1 occlusive stroke admitted to the Department of Neurology, the Second Affiliated Hospital of Anhui Medical University from June 2018 to September 2019 were enrolled retrospectively. The demographic and clinical data were collected. Diffusion-weighted imaging (DWI)-Alberta Stroke Program Early CT Score (ASPECTS) and FVH score were performed with MRI images. The modified Rankin Scale (MRS) was used to evaluate the clinical outcome at 90 d after onset. 0-2 was defined as good outcome, and >2 was defined as poor outcome. Multivariate logistic regression analysis was used to determine the independent correlation between FVH and the outcome. Results:A total of 65 patients with acute middle cerebral artery M1 occlusive stroke were enrolled, including 37 males (56.9%). Their age was 64.35±12.13 years. Twenty-nine patients (44.6%) had a good outcome, and 36 (55.4%) had a poor outcome. There were significant differences in triglyceride ( P=0.037), antihypertensive drug treatment ( P=0.037), baseline National Institutes of Health Stroke Scale (NIHSS) score ( P<0.001), DWI-ASPECTS ( P=0.017) and FVH score ( P<0.001) between the poor outcome group and the good outcome group. Multivariate logistic regression analysis showed that FVH score (odds ratio 6.477, 95% confidence interval 1.570-26.716; P=0.010) and NIHSS score (odds ratio 1.869, 95% confidence interval 1.326-2.635; P<0.001) were significantly independently correlated with the poor outcome. However, there was no significant independent correlation between DWI-ASPECTS and the outcome (odds ratio 0.451, 95% confidence interval 0.068-2.988; P=0.410). Conclusions:FVH score is an independent risk factor for poor outcome in patients with acute middle cerebral artery M1 occlusive stroke.

OBJECTIVETo investigate the correlation between miR-221 and epithelial-mesenchymal transition (EMT) in drug-resistant glioma cells.

METHODSThe expression levels of miR-221, PTEN, p-Akt, E-cadherin, vimentin, and MRP1 were quantitatively analyzed in Z1 cells (primary drug-resistant cells), Z2 cells (drug-sensitive cells) and Z2-BCNU cells (drug-resistant cells) using fluorescent real-time PCR and Western blotting.

RESULTSThe expression levels of PTEN were significantly increased in Z2 cells compared with Z1 and Z2-BCNU cells which overexpressed miR-221 and vimentin. The expression levels of vimentin, p-Akt and MRP1 were significantly decreased in Z2 cells overexpressing E-cadherin.

CONCLUSIONMiR-221 regulates the expression of EMT-related genes through down-regulation of PTEN and activation of PI3-K/Akt signaling.

Cadherins ; metabolism ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Epithelial-Mesenchymal Transition ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; genetics ; metabolism ; Multidrug Resistance-Associated Proteins ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Vimentin ; metabolism

AIM: To study the pathological change in mouse organs immunitied by inactivated SARS-CoV vaccine. METHODS: Inactivated SARS-CoV vaccine was injected into BALB/c and C57BL/6 mice. Anti-SARS antibody was analyzed by ELISA. After 8 weeks, the immunitied mice were killed and those organs were analyzed by pathological methods. RESULTS: Anti-SARS antibody in mice was positive after 8 days. Only minimal injury was observed in a few lungs and livers, but the other organs were not. CONCLUSIONS: Inactivated SARS-CoV vaccine induced mice to create antibody, whereas they did not cause severe injury. This result will be valuable for vaccine into clinical research. [