Objective To evaluate an ideal way to prepare the extracellular matrix of urethra. Methods An orthogonal design [L9(34)] was used in the experiment.Urethras were obtained from 37 rabbits,among which 27 segments were randomly selected and were decellularized following the orthogonal design in 9 groups.The whole experiments were repeated for 3 times.After the decellularization process,the acellularity of the ECM was examined by haematoxylin-eosin staining.The optimum way was found out through comparing the numbers of the remained cellular elements by computer image analysis.An ideal way was found by statistic analysis.Then the ECM was obtained from 10 pieces of urethras by the optimum methods.The scanning electronic microscopy was used to confirm the decellulary matrix.Subsequently,the ECM was used as a graft for replacement. In 10 rabbits,the urethral defect were replaced with the urethral ECMs. At sacrifice,10 days,3 weeks,6 weeks and 24 weeks,the grafts was taken out,and the regeneration was confirmed by the haematoxylin-eosin staining. Results ECM resulting from different dedellularization process in the urethras are different in the numbers of remaining cellular elements.There are no cellular elements in the 7th and the 9th group of the tissues.The cellular elements was not found by the scanning electronic microscopy in the ECM getting from the optimum methods.In the animals with replacement,histologic examination showed complete regeneration 24 weeks post operation. Conclusions The best way to prepare the ECM of urethra is A 3B 2C 3.

Objective To investigate the clinical application of parenteral nutrition drugs and promote the rational use of drugs.Methods The data of parenteral nutrition drugs used in our hospital were analyzed,including the quantity,the amount of money,the scope and purpose,etc.The rationality of drug use was reviewed by prescription comment simultaneously.Results On similar medicines comparison,the fat emulsion,amino acids (17) and glucose (11％) injection (1440mL) and fat emulsion and amino acids (18) injection (1 000mL),which were designed according to the basal metabolic rate in patients were far greater than the fat emulsion,amino acids(17) and glucose (11％) injection (1 920mL) of the added value of design.And the balanced amino acid preparations in compound amino acid in 18 AA-Ⅳ usage,was far greater than 18 AA-Ⅱ and 18 AA.Fat emulsion preparation with concentration of 20％ dosage was bigger,because the price advantage,ordinary long-chain fat milk consumption was greater than the long chain fatty milk.In terms of drug use rationality,the usage,dosage,compatibility and adverse reactions of parenteral nutrition drugs were analyzed.The dosage,frequency and compatibility problems still needed to be improved.Conclusion The use of parenteral nutrition drugs were basic reasonable,but we should strictly control the indications for parenteral nutrition support,such as drug compatibility,using frequency to strengthen management and reasonable application of drugs to improve medication safety.

Objective To evaluate if the flexible ureteroscopy could treat stones located in lower calyx with the infundibulopelvic angle (IPA) less than 30°.Methods Thirty-six patients with inferior caliceal calculi on whom flexible ureteroscopic procedures were performed between November 2009 and June 2015 were reviewed.The mean age of the patients was 52.1 years (34-71),with the mean stone diameter of (1.5 ± 0.8) cm (1.2-2.6 cm).IPA was smaller than 30° in all 36 cases,which confirmed by CTU examination.IPA was less than 10° in 15 patients,between 11 ° and 20° in 13 patients and between 21 ° and 30° in 8 patients.Results The success rate was 63.9％ (23/36 patients) in patients with IPA smaller than 30° after first session of procedure,and the stone free rate reached 100％ after the second session of procedure.The mean operation duration was (95.5 ± 31.4) min(51-127 min).The mean hospital stay after operation was(4.1 ± 1.2)days (3-5 days).No major complications were recorded and no patients needed to convert to open surgery.Double J tube was removed after 4 weeks postoperatively.Patients were followed up for 4-12 months,during which ultrasound and CT scan were used for stone detection.Conclusions The small IPA (＜30°) negatively affected the SFR in the first session operation.However,its negative effect was solvable by using modern endoscopes.A complete stone clearance was achievable even in case of unfavorable anatomic conditions in experienced hand.

BACKGROUND:The main way for long-segmental ureteral reconstruction may cause a lot of traumas and complications. Therefore, to seek a new repair method is urgent. OBJECTIVE:To investigate the feasibility of a tissue-engineered tubular graft for ureteral reconstruction. METHODS:Bone marrow mesenchymal stem cels and smooth muscle cels of rabbits were seeded into the two surfaces of bladder acelular matrix and cultivated for 7 days. Then the graft was used to prepare a 4-cm long tissue-engineered tubular graft, which was regarded as experimental group. Smooth muscle cels seeded onto the bladder acelular matrix was used to construct the tissue-engineered tubular graft as control group. Twenty-five New Zealand rabbits were randomly divided into experimental group (n=20) and control group (n=5), and two kinds of tubular grafts covered with omentum were implanted into the two groups, respectively, for repair of ureteral defects. Hematoxylin-eosin staining and immunohistochemical detection were performed at 2, 4, 8 weeks after implantation. RESULTS AND CONCLUSION:In the experimental group, hematoxylin-eosin staining showed epithelial coverage and muscle fibers on the lumen of tissue-engineered tubular grafts at 8 weeks after implantation; immunohistochemistry showed that anti-AE1/AE3 antibody and anti-uroplakinⅢa antibody were positive, confirming that there were mature epithelial cels on the lumen of tissue-engineered tubular grafts. In the control group, five rabbits were dead within 2 weeks after removal of ureteral scaffold, and autopsy showed scar formation inside the graft and severe hydronephrosis. These results demonstrate that it is feasible to construct the tissue-engineered tubular graft using bone marrow mesenchymal stem cels and smooth muscle cels into the bladder acelular matrix for ureteral reconstruction. Bone marrow mesenchymal stem cels can potentialy promote urothelial regeneration.

BACKGROUND:Urothelial cells are important seeding cells for urinary tissue engineering, but they are difficult to proliferate in vitro. Several studies have shown that bone marrow mesenchymal stem cells can differentiate into urothelial cells, but how these cells functions in vivo in epithelium generation after implantation, and the application of these cells in tissue engineering, are rarely studied. OBJECTIVE:To explore the isolation and proliferation of rabbit bone marrow mesenchymal stem cells that are induced into urothelial cells in combination with rabbit bladder acellular matrix to construct tissue-engineered grafts, and to assess the effect of the induced cells as seeding cells. METHODS:Twelve 8-week-old male New Zealand white rabbits were chosen to obtain bone marrow samples through tibia puncture, and to isolate bone marrow mesenchymal stem cells by density gradient centrifugation. Then the fourth or fifth generation of bone marrow mesenchymal stem cells were cultured in conditioned medium for 2 weeks, and then identified by PCR and immunofluorescence. After that, the induced cells were seeded on rabbit bladder acellular matrix to construct tissue-engineered grafts for bladder repairing. Another 12 rabbits served as control group, and urothelial cells combined with bladder acellular matrix was used for bladder repairing. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells were successful y cultured and proliferated in vitro. After induction, PCR detection suggested that stem cellmarker (CD44) expression decreased, and epithelial cellmarker (UP1a) expression increased in the induced cells. Immunofluorescence staining demonstrated that the induced cells rather than bone marrow mesenchymal stem cells were positive for specific urothelial marker, UP1a. A stable continuous epithelial layer was observed on tissue-engineered grafts constructed by induced cells after 2 weeks, similar to the grafts built by urothelial cells. Induced bone marrow mesenchymal stem cells can differentiate into urothelial cells that can be used as seeding cells for urinary tissue engineering, which may be another choice out of urothelial cells.

Objective To explore the effect of Yiqi-jianpi-jiedu recipe decoction containing serum on Oxaliplatin-induced HepG2 Human hepatoma cell survivin, caspase-3, caspase-9 expression. Methods The serum pharmacological method was used to prepare nourishing Qi and invigorating spleen decoction containing serum of rat. The HepG2 human hepatoma cells were divided into Chinese herb containing serum group, chemotherapy with Oxaliplatin (OXA) group and the combination group (medicine containing serum and OXA), and the blank group without drugs. Apply MTT assay, Western Blotting method and RT-PCR method to detect the cell inhibition rate and survivin, caspase-3, caspase-9 protein and gene expression changes. Results Compared with the blank group, the inhibition rate of HepG2 Human hepatoma cell was significantly higher in Chinese herb containing serum group, chemotherapy group and the combination group (P < 0.05). Compared with thechemotherapy group and Chinese herb containing serum group, the Survivin (0.151 ± 0.054 vs. 0.288 ± 0.089, 0.375 ± 0.063) and the expression of Survivin mRNA (0.205 ± 0.091 vs. 0.487 ± 0.073, 0.725 ± 0.092) of the conbination group downgraded significantly lower (P < 0.05 or P< 0.01). Compared with chemotherapy group and Chinese herb containing serum group, the caspase-3 (0.821 ± 0.079 vs. 0.634 ± 0.098, 0.487 ± 0.102), caspase-9 (0.901 ± 0.047 vs. 0.709 ± 0.054, 0.402 ± 0.012) expression, caspase-3 mRNA (1.928 ± 0.226 vs. 1.564 ± 0.195, 1.287 ± 0.312) and caspase-9 mRNA (2.063 ± 0.517 vs. 1.536 ± 0.084, 1.019 ± 0.182) expression of the conbination group upgraded significantly higher (P < 0.05 or P < 0.01). Conclusions Nourishing-Qi and invigorating spleen decoction containing serum may affect Oxaliplatin-induced HepG2 apoptosis mechanism through increasing the suppression on Survivin and promoting the expression of caspase-3 and caspase-9 protein and genes. This research provided important experimental basis for clinical application of traditional Chinese medicine treating liver cancer.

OBJECTIVE:To standardize the clinical use of adjuvant therapy drugs. METHODS:To establish the clinical use management mode of adjuvant therapy drugs in our hospital and summarize the usage amount and irrational drug use of adjuvant therapy drugs before and after implementing the management,and the management effect was assessed. RESULTS:The manage-ment mode of adjuvant treatment drugs in our hospital included ensuring the directory for adjuvant therapy drugs,establishing the management system and ensuring its basic principles,then conducting drug use review with the ranking of drug dosages and mak-ing full use of evidence-based medical evidence,as well as developing principle of classification disposal for the irrational drug use (off-indications,overdose,ultra treatment) of adjuvant therapy drugs. After more than one year of management practice,the use of adjuvant therapy drugs in our hospital is increasingly standardized. The sales amount of adjuvant therapy drugs to all drugs was reduced from 20.90％ before management(Aug. 2014-Jul. 2015)to 15.99％(reduced by 5％)after management(Aug. 2015-Jul. 2016). Compared with Aug. 2015,the sales amount of 48 varieties in the 93 adjuvant therapy drugs was dropped more than 15％in Aug. 2016,and irrational drug use decreased obviously. CONCLUSIONS:The established management mode of adjuvant thera-py drugs in our hospital has effectively improved clinical rational drug use and reduced medical costs.

0.05).Conclusion: The method of constructing a highly invasive subline of bladder neoplasm is reliable,and EGFP transfection does not change the biological behavior of the cells.The obtained sublines may provide a valuable experimental platform for further study on the molecular mechanisms of bladder neoplasm metastasis.

This study examined the osteogenic effect of electromagnetic fields (EMF) under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells (BMSCs) and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit (CCK-8). For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase (ALP) staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure (2 h/day) could obviously promote proliferation of BMSCs and osteoblasts, while long-term EMF (8 h/day) could promote osteogenic differentiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when cells were exposed to EMF. Compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differentiation of BMSCs.

A disintegrin-like and metalloprotease(ADAMTS) proteinases are a group of multidomains and secreted metalloproteinases containing the thrombospondin motifs. ADAMTS-7 is a member of ADAMTS family and plays a crucial role in the cardiovascular diseases. In physically, ADAMTS-7 was involved in the growth and development of the body. However, in pathological conditions, over expression of ADAMTS-7 gene accelerated the progression of cardiovascular diseases through by promoting the breakdown of cartilage oligomeric matrix protein (COMP) matrix. Meantime, the over expression of ADAMTS-7 can be effectively inhibited by TGF-β, miRNA29a/b, and then effectively retarded the development of cardiovascular disease. This article mainly reviews the gene structure, protein function and the mechanism of ADAMTS-7 involved in cardiovascular diseases and looks forward to the possibility of ADAMTS-7 as a new target for the treatment of cardiovascular diseases.